Genetically engineering cotton plants for altered fiber

ABSTRACT

A method of genetically engineering a fiber-producing plant is disclosed. This method first comprises the step of constructing a plant expression vector that comprises a protein coding sequence and a DNA sequence capable of promoting gene expression in fiber cells, wherein the DNA sequence is homologous to a sequence selected from the group consisting of 12 different genomic sequences. The method next involves introducing the expression vector into a fiber-producing plant wherein the protein coding sequence is expressed in the fiber cells of the fiber producing plant. A method of obtaining a DNA sequence capable of promoting gene expression in fiber cells is also disclosed.

CROSS REFERENCE TO RELATED APPLICATIONS

This is a division of application Ser. No. 07/885,970, filed May 18,1992, which is a continuation-in-part of Ser. No. 07/617,239, filed Nov.21, 1990 (now abandoned), which is a continuation-in-part of Ser. No.07/253,243 filed Oct. 4, 1988 (now abandoned). Both of theseapplications are hereby incorporated by reference.

FIELD OF THE INVENTION

The present invention relates to the general technology of plant geneticengineering and, in particular, to the identification of fiber-specificpromoters and the use of these promoters to create novel geneticallytransformed cotton (Gossypium) plants and lines with varied cotton fibercharacteristics and quality.

BACKGROUND OF THE INVENTION

Genetic Engineering of Plants

The hurdle of creating successful genetically engineered plants in majorcrop varieties is now being overcome sequentially on a plant-by-plantbasis. While plant genetic engineering has been successfullydemonstrated in several model plant species, most notably tobacco,carrot and petunia, these species are not considered agriculturallyimportant. Therefore, researchers have now directed their efforts towardimproving commercially important crop plants through the use of geneticengineering (Potrykus, I., Annu. Rev. Plant. Physiol. Mol. Biol.42:205-225, 1991).

The term "genetic engineering," as used herein, is meant to describe themanipulation of the genome of a plant, typically by the introduction ofa foreign gene into the plant, or the modification of the genes of theplant, to increase or decrease the synthesis of gene products in theplant. Typically, genes are introduced into one or more plant cellswhich can be cultured into whole, sexually competent, viable plantswhich may be totally transformed or which may be chimeric, having sometissues transformed and some not. These plants can be self-pollinated orcross-pollinated with other plants of the same or compatible species sothat the foreign gene or genes carried in the germ line can be bred intoagriculturally useful plant varieties.

Current strategies directed toward the genetic engineering of plantlines typically involve two complementary processes. The first processinvolves the genetic transformation of one or more plant cells of aspecifically characterized type. The term "transformation" as usedherein means that a foreign gene, typically in the form of a geneticconstruction, is introduced into the genome of the individual plantcells. This introduction is typically through the aid of a vector, whichis integrated into the genome of the plant. The second process theninvolves the regeneration of the transformed plant cells into wholesexually competent plants. Neither the transformation nor regenerationprocess need be 100% successful, but must have a reasonable degree ofreliability and reproducibility so that a reasonable percentage of thecells can be transformed and regenerated into whole plants.

Genetic Engineering of Cotton

Although successful transformation and regeneration techniques have beendemonstrated in model plant species. (Barton et al., Cell 32:1033(1983), wherein the transformation and regeneration of tobacco plantswas reported) similar results with cotton have only been achievedrelatively recently. Umbeck et al. Bio/Technology, 5[3] 263-266 (1987);Firoozabady et al., Plant Mol. Bio. 10:105-116 (1987); Finer et al.,Plant Cell Rep. 8:586-589, 1990.

Successful transformation and regeneration of genetically engineeredcotton plants has the potential to be of significant value to thisagriculturally important crop. One of the most important benefitspotentially achievable from genetically engineering cotton plants is thealteration and modification of cotton fiber quantity and quality.

Cotton Fiber

Cotton is one of the most important cash crops. Cotton fiber (seed hair)is a differentiated single epidermal cell of the ovule. At maturity thefiber cell consists of a cell lumen, primary cell-wall and secondarycell-wall. The primary cell-wall is made up of pectic compounds,cellulose, and small amounts of protein. The secondary cell-wallconsists of cellulose. At maturity, the cotton fiber contains 87%cellulose.

Cotton fiber development can be divided into initiation, primarycell-wall synthesis stage, secondary cell-wall deposition stage, andmaturation phases. Many hundreds of genes are required for thedifferentiation and development of cotton fiber. Work on in vitrotranslated fiber proteins (Delmer et al., J. Cell Sci. Suppl. 2:33-50,1985), and protein isolated from fiber (Graves and Stewart, J. Exp. Bot.39:59-69, 1988) clearly suggests differential gene expression duringvarious developmental stages of the cell. However, none of the genesinvolved in the biosynthesis of the large numbers of fiber-specificstructural proteins, enzymes, polysaccharides, waxes or lignins havebeen identified. Since these genes and their interactions withenvironment determine the quality of fiber, their identification andcharacterization is considered to be an important aspect of cotton cropimprovement. The current invention is designed to approach fibermodification through genetic engineering. Such an endeavor requiresfiber-specific promoters, genes that will modify fiber properties, andan efficient transformation technique.

The quality of the cotton fiber is dependent on such factors as theextent of elongation and degree of secondary wall deposition. It isassumed that a number of genes as well as environmental factors regulatethe physical characteristics of the fiber, such as length, thickness andmicronaire value. However, the genes responsible for cellulose synthesisand fiber development in cotton plants are heretofore entirelyuncharacterized at a molecular level.

The most commercially useful plant fiber is derived from cotton(Gossypium arboreum, Gossypium herbaceum, Gossypium barbadense andGossypium hirsutum). However, there are other fiber-producing plants.These plants include the silk cotton tree (Kapok, Ceiba pentandra),desert willow, creosote bush, winterfat, balsa, ramie, kenaf, hemp,roselle, jute, sisal abaca and flax.

Promoters

Promoters are DNA elements that direct the transcription of RNA incells. Together with other regulatory elements that specify tissue andtemporal specificity of gene expression, promoters control thedevelopment of organisms. Thus, there has been a concerted effort inidentifying and isolating promoters from a wide variety of plants andanimals.

Many promoters function properly in heterologous systems. For example,plant gene promoters such as rbcS, Cab, chalcone synthase and proteaseinhibitor from tobacco and Arabidopsis are functional in heterologoustransgenic plants. (Reviewed by Denfey, et al. Science 244:174-181,1989). Specific examples of transgenic plants include tissue-specificand developmentally regulated expression of soybean 7s seed storageprotein gene in transgenic tobacco plants (Chen, et al. EMBO J.7:297-302, 1988.) and light-dependent organ-specific expression ofArabidopsis thaliana chlorophyll a/b binding protein gene promoter intransgenic tobacco (Ha and An, Proc. Nat'l. Acad. Sci. USA 85:8017-8021,1988). Similarly, anaerobically inducible maize sucrose synthase-1promoter activity was demonstrated in transgenic tobacco (Yang andRussell, Proc. Nat'l. Acad. Sci USA, 87:4144-4148, 1990). Tomato pollenpromoters were found to direct tissue-specific and developmentallyregulated gene expression in transgenic Arabidopsis and tobacco (Twellet al., Development 109:705-713, 1990). Thus, some plant promoters canbe utilized to express foreign proteins in plant tissues in adevelopmentally regulated fashion.

SUMMARY OF THE INVENTION

The present invention is a method of creating a transgenicfiber-producing plant. This method comprises the steps of constructing aplant expression vector that comprises a protein-coding sequence and apromoter DNA sequence and introducing the expression vector into afiber-producing plant. The protein coding sequence is expressed in thefiber cells of the fiber-producing plant. By "protein coding sequence"we mean a sequence that encodes at least a portion of a protein and isin either the sense or antisense orientation. By "expressed" we mean toinclude sequences expressed as RNA and as protein. Preferably, the DNAsequence is homologous to a sequence selected from the group consistingof the gene E6 4.5 Kb Mbo I/Nco I fragment; the gene E6 2.7 Kb Mbo I/NcoI fragment; the gene E6 4.1 Kb Nco I fragment; the gene E6 3.9 Kb MboI/Nco I fragment; the gene E6 3.2 Kb Mbo I/Nco I fragment; the gene H6321 bp Fsp I/Sal I fragment; the gene B6 2.3 Kb Bam HI/Xho I fragment;the gene E9 7.0 Kb Xho I/Nco I fragment; the gene A12 3.5 Kb Mbo I/Sma Ifragment; the gene A-11 2.1 Kb Mbo I/Eco RI fragment; the gene B8 2.2 KbBam HI/Bst BI fragment; and the gene B12 3.1 Kb Eco RI/Sty I fragment.The homology to the sequence is sufficient to provide promoter activity.

Also preferably, the DNA sequence is homologous to a sequence selectedfrom the group consisting of SEQ ID NOs: 20-29. The homology issufficient to provide promoter activity.

The present invention is also a method of obtaining a DNA sequencecapable of promoting gene expression in fiber cells. The method involvesfirst obtaining a genomic library prepared from the DNA of afiber-producing plant. This genomic library is screened with anucleotide sequence that is homologous to an RNA sequence preferentiallyexpressed in fiber cells or with a nucleotide sequence from a genomicclone homologous to a fiber-specific RNA. A homologous genomic sequenceis selected by this screening. A promoter sequence is isolated from thegenomic sequence. Preferentially, the genomic library is screened with anucleotide sequence sufficiently homologous to a sequence selected fromthe group consisting of SEQ ID NOs: 2-29 to hybridize to a genomicsequence with promoter activity.

It is an object of the present invention to genetically engineer cottonplants and lines and other fiber-producing plants and lines.

It is another object of the present invention to genetically engineerfiber-producing plants in order to alter fiber quantity and quality.

It is another object of the present invention to identify promoterswhich regulate fiber production or fiber-specific genes in cotton orother fiber-producing plants.

It is another object of the present invention to express foreign genesin a plant in a fiber-specific manner.

It is a feature of promoter DNA sequences identified by the presentinvention that these promoters may be identified in one plant yet usefulin another fiber-producing plant.

It is another feature of the promoter DNA sequences identified by thepresent invention that they contain DNA sequences sufficient to directtranscription and that they contain DNA sequences sufficient fortissue-specific transcription.

It is an advantage of the present invention that genetically-engineeredcotton plants with altered fiber quantity or quality may be able toproduce progeny with this trait.

Other objects, features and advantages of the present invention willbecome apparent upon examination of the specification, drawings, andclaims.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A and 1B are a set of restriction maps of cotton genomicfragments assayed for promoter activity.

FIG. 2 is a bar graph describing the levels of promoter activityobtained in a fluorogenic assay with the fragments of FIG. 1.

FIG. 3 is a diagram of plasmid p2117.

FIG. 4 is a diagram of the construction of plasmid p2117E6P-2A.

FIG. 5 is a diagram of the construction of plasmid p2117E6P-3B.

FIG. 6 is a diagram of plasmid pCPE6-2117.

FIG. 7 is a diagram of H6 cDNA.

FIG. 8 is a diagram of the H6 gene.

FIG. 9 is a diagram of plasmid pH6-2117.

FIG. 10 is a diagram of the construction of plasmid pE6-3B-ExY.

DETAILED DESCRIPTION OF THE INVENTION

These objects and others are fulfilled by the present invention whichinvolves a method of creating transgenic fiber-producing plants. In manyinstances it is desirable for the transgene to be developmentallyregulated so as to be expressed only in fiber cells at a properdevelopmental stage. This regulation can be most expeditiouslyaccomplished by promoters capable of preferential promotion. Thesepromoters may be obtained by using cDNA clones from fiber-specific mRNAsto find genomic clones from a genomic library of the plant. From thegenomic clone, the entire fiber-specific gene, including developmentallyregulated promoter and regulatory sequences, can be isolated.

Therefore, the method first involves identifying promoters thatpreferentially promote gene expression in fiber cells. (By"preferentially promote" we mean that the gene is either expressed onlyin fiber cells or is expressed in fiber cells more actively than it isexpressed in other plant tissue cells.) One way to identify suitablepromoters is by isolating mRNA from fiber-producing cells, making acomplementary DNA (cDNA) library of cDNA clones from the mRNA, andscreening the cDNA library with cDNA generated from other tissues toidentify and eliminate RNAs which are expressed in tissues other thanthose which produce fiber. This screening procedure will result in theidentification of cDNA clones that are expressed preferentially in fibercells. The sequences of eighteen cDNA clones from fiber-specific mRNAsare given below at SEQ ID NOs: 2-19.

After the identification process is complete, fiber-specific cDNA clonesmay be used to screen genomic clones created from the DNA offiber-producing plants. This screening process results in the selectionof genomic clones with sequences homologous to the fiber-specific cDNAs.Because a fiber-specific promoter will be upstream from a sequence thatis expressed specifically in a fiber cell, the promoter sequence may beidentified on this genomic clone. These promoter sequences may beexcised and attached to genes which if expressed in fiber cells wouldalter fiber quality or quantity. (Although the plant may be any of anumber of varieties of fiber-producing plants, cotton (Gossypium) plantsare the preferred plants for purpose of the present invention).

The present invention is preferably performed with one of twelve cottongenomic DNA fragments that we have identified as containing promoteractivity. SEQ ID NOs: 20-29 represent the DNA sequences of ten of thesefragments. Some of the ten fragments have only been partially sequenced.Two of the fragments, the gene E9 7.0 Kb Xho I/Nco I fragment and thegene A12 3.5 Kb Mbo I/Sma I fragment, have not been sequenced.

A sequence that is only a portion of one of these twelve identifiedfragments may also contain promoter activity because it is not necessaryfor a DNA fragment to contain an identical nucleotide sequence to befunctionally identical to the promoter sequences described here. Thesequence must only be sufficiently homologous to the fragment to retainpromoter activity. Some nucleotide deletions, additions, andreplacements, either naturally occurring or artificially induced, willhave only a minor impact on gene expression.

The twelve promoter fragments may be truncated to determine the smallestfragment capable of tissue-specific expression. Methods of truncating aclone include deleting sequences and digesting the clone with arestriction enzyme or other nuclease. These methods are commonly knownin the art of molecular biology. The promoter assay described below willenable one to determine whether or not a specific portion of DNAcontains promoter activity. Creation of a transgenic plant enables oneto determine whether a DNA fragment contains a sequence sufficient fortissue specificity.

Another way to obtain a sequence capable of preferentially promotingexpression in fiber-producing plants is to probe a library of DNAobtained from a fiber-producing plant with a probe prepared from SEQ IDNOs: 2-19 (the fiber-specific cDNAs) or SEQ ID NOs: 20-29 (the genomicfragments). The DNA probe must only be of a length sufficient tohybridize specifically to a suitable clone. If a sequence from afiber-specific cDNA clone is used, one will isolate the genomic clonehomologous to that cDNA. Because promoter sequences are found upstreamfrom the sequences homologous to the cDNA clone, one must examine theseupstream sequences to find the promoter.

If one wishes to recreate the twelve fragments we have assayed forpromoter activity, one would first screen a cotton genomic library witha probe prepared from SEQ ID NOs: 2-29. For example, if one wished torecreate the gene B6 2.3 Kb Bam HI/Xho I fragment, one would screen thegenomic library with a probe prepared from SEQ ID NO: 29 (the genomicfragment) or SEQ ID NO: 15 (B6 cDNA). The identified genomic clone wouldbe subjected to restriction digests with Bam H1 and Xho I to locate the2.3 Kb fragment.

After a fiber-specific promoter has been identified and isolated, thepromoter must be placed upstream of a gene whose expression is desired.Preferably, the product of this gene is capable of altering fiberquality or quantity. Conventional molecular biological techniques may beused to create suitable constructs.

These constructs must be transformed into a cotton plant or cell. Stableintegration and expression of foreign genes in cotton plants has beendemonstrated and repeated. Umbeck et al., Bio/Technology, 5[3]:263-266(1987); Firoozabady et al., Plant Mol. Biol., 10:105-116 (1987). Usingthe techniques taught in these papers, the transformation of cottontissues is accomplished by Agrobacterium infection and regeneration.Although a lengthy process, the Agrobacterium-mediated transformation ofcotton has also been practiced by other laboratories and can now readilybe replicated by those of ordinary skill in plant genetic engineering.

It is to be understood, however, that other methods for thetransformation of cotton plants and lines are being studied, and thatthe transgenic cotton plants and lines with fiber genes introduced intothem will prove advantageous and useful regardless of the method oftransformation of the original tissues. Specifically, it has now beendemonstrated that higher plants can be stably genetically transformed byparticle-mediated transformation techniques, which avoid many of thedifficulties and delays inherent in plant regeneration required byAgrobacterium plant transformation. McCabe et al., Bio/Technology,6[8]:923-926 (1988). Recent research results suggest that routineparticle-mediated transformation of cotton is to be expected shortly.

The present invention is a useful genetic engineering tool for theintroduction of altered fiber-specific characteristics into cottonplants. The identification and introduction of fiber-specific promotersfrom one plant variety to another can be extended to include otherexotic plants that produce fiber. Many of these plants will havefiber-specific promoters with one or more desirable qualities, which canbe incorporated into a cotton plant.

The promoters of the present invention can be utilized in modulating thesynthesis of fiber proteins or to introduce non-fiber proteins intofiber in a tissue-specific manner. To determine the sequences within agene necessary for fiber-specific expression, nucleotide sequences ofthe coding region and regions flanking the coding region can besubjected to computer analysis to identify sequence patterns thatcorrespond to consensus regulatory elements. Potential regulatoryelements are usually present at the 5' flanking region of the gene, 30to 100 bases upstream from the transcription start site in eukaryoticgenes (for reviews see Breathnach and Chambon, Annu. Rev. Biochem.50:349-383, 1981; Johnson and McKnight, Annu. Rev. Biochem. 58:799-839,1989). In addition to the promoter (TATA box) other consensus sequencessuch as the CATC box and the CACA box, may also be present in specificgroups of genes. Messing, J. et al., in Genetic Engineering of Plants,an Agricultural Perspective, Kosuge, et al. eds. pp 211-227 (1983);Forde, B. G. et al., Nucl. Acid Res. 13:7327-7339 (1985); Goldberg, R.B. Philos. Trans. Roy Sci. B314:343-353 (1986). A search of the 5'flanking sequences of the gene can identify these sequence patterns.

The presence of these consensus sequences in the 5' region is anindication of promoter activity, but the presence of these consensussequences is not conclusive that the identified DNA segment is the truepromoter. This is because many concensus sequences can be located in agiven gene. Therefore, true identity of the promoter element requiresother promoter assays.

One may use a transient reporter gene expression system to assesspromoter activity. In such an assay, the fragment to be assayed would belinked to a reporter gene and used to transform a plant cell. Usefulreporter genes include chloramphenicol acetyltransferase (CAT),luciferase (Lux) and β-glucuronidase (GUS). Alam and Cook, Anal.Biochem. 188:245-254, 1990; Jefferson, Plant Mol. Biol. Rep. 5:387-405,1987. We have described a reporter gene assay in our examples. Furtherconfirmation of the promoter activity and tissue-specific anddevelopmental expression can be obtained by stably integrating achimeric construct comprised of the DNA segment and reporter gene intoplants or animals and following the reporter gene's expression throughdevelopment.

Another approach to creating cotton plants with altered fibercharacteristics is to create antisense genetic constructs withfiber-specific promoters to inhibit or lessen the expression of one ormore fiber genes in fiber cells. The theory behind antisense geneticconstructs is that the production of RNA strands in the cells of anorganism which are complementary to the mRNA of an endogenous gene willresult in hybridization of the antisense RNA to the native mRNAresulting in decreased expression of the mRNA gene. Smith, et al. Nature334 724-726, 1988; Bird, et al. Bio/Technology 9:635-639, 1991; Van derKrol, et al. Gene 72:45-50, 1988. Thus, in an antisense construct, acomplete coding sequence for the mRNA is not needed. All that is neededis a sequence of sufficient length to construct a selectivelyhybridizing antisense RNA. Thus, the cDNA clones discussed below are ofparticular utility for this approach.

The following is a description of the process and materials used toidentify fiber-specific promoters and to transform cotton plants.Although reference to cotton is specifically made, it is within thescope of the present invention to substitute other fiber-producingplants.

EXAMPLES

1. Isolation of RNA From Fiber

Fiber cells at different stages of development from fiber-producingplants were collected and quick-frozen in liquid nitrogen. Specifically,fiber cells from 15 and 23 day-old Coker 312 or 10 day-old Sea Islandbolls were collected and quick-frozen. The frozen fiber cells were thenpowdered in a mortar in liquid nitrogen and homogenized in ahomogenization buffer for 1.5 minutes using a polytron at full speed.The homogenization buffer included the following ingredients: 5 MGuanidine isothiocyanate; 0.2 M Tris-acetate (pH 8.5); 0.7%Beta-mercaptoethanol; 1% polyvinyl pyrrolidone (PVP, MW 40 Kd), and0.62% sodium lauroyl sarcosine. Beta-mercaptoethanol and PVP were addedjust before use. A ratio of 1:2 of tissue (weight) to buffer (volume)was used.

The homogenate was filtered through Mira cloth and layered over a 1.5 mlpad of 5.7 M cesium chloride as described by Chirgwin, J. M. et al.Biochemistry, 18: 5294-5299 (1979). The homogenate was then centrifugedfor 18 hours at 36,000 rpm in a SW 50.1 rotor at 20° C. Aftercentrifugation, the RNA was collected as described by Chirgwin et al.(supra).

The RNA was then further purified by phenol:chloroform extractions andprecipitations in the presence of ammonium acetate as described for DNAby Crouse, et al., Focus, 9[2]:3-5 (1987). Poly(A)+ RNA was obtained byoligo-(dT) chromatography as described by Maniatis, et al., in MolecularCloning: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold SpringHarbor, N.Y., (1982).

2. Library Construction and cDNA Clone Identification

Complementary DNA (cDNA) libraries were prepared from the mRNA accordingto the protocol developed by D'Alessio et al., Focus, 9[1]:1-4 (1987)with the following exceptions: The first strand of cDNA was synthesizedusing a primer having the following sequence dATGCTGGTACC(T)₅ (SEQ IDNO: 1); the second strand synthesis was carried out as described byD'Alessio et al., supra, for tailing. The poly-(dC) tails were added tothe double-stranded cDNA and then annealed to poly-(dG)-tailed pBR322plasmid vector (Bethesda Research Laboratories). The recombinantplasmids were used to transform Escherichia coli (E. coli) RR1 strain asdescribed by Hanahan in DNA Cloning a Practical Approach, Vol. 1 (1985)p. 109-135. The transformed cells were selected on agar platescontaining the antibiotic tetracycline (12 mg/liter).

Separate cDNA libraries were constructed from the mRNAs from 10-day,15-day, and 23-day-old fiber cells. For the 10-day fiber cell mRNAs, anoligo-(dT) primer was used for cDNA synthesis instead of the primerdescribed above. The 10-day cells were selected to be representative ofgenes active during the primary cell wall stage of cell development. Inthe 15-day-old cell, both primary cell wall and secondary cell wallsynthesis systems are active. The 23-day-old cells were selected to berepresentative of genes active principally during secondary wallsynthesis.

The clones in the library were then transferred to nitrocellulosefilters and duplicate filters were made according to Hanahan, et al.,Gene, 10:63-67 (1980). About 25,000 clones from the 15-day and 23-daylibraries were screened using the following procedure. ³² P-labelledsingle-stranded cDNA probes were prepared from poly(A)+ RNAs using ³²P-dCTP and reverse transcriptase as described by Maniatis et al., supra.Probes were prepared from poly(A)+ RNAs of 15-day, 23-day old fiberproducing cells, and from 0-day ovule, leaf, root and flower cells.Prewashings, prehybridizations, hybridizations and washings of thefilters were performed as described in detail in John et al., Proc.Natl. Acad. Sci. USA, 81:5628-5632 (1984).

The autographic signals from filters hybridized with ³² P-labelled cDNAsfrom the different tissues were then compared. The clones whichhybridized to cDNAs from fiber producing cells, but not to cDNAs fromother tissues, were selected. The resulting clones were then subjectedto a second cycle of differential screening as described above andadditional clones were eliminated as non-fiber specific. This processwas continued for a third and then a fourth time. This repetitivescreening was to eliminate clones which showed hybridization to otherthan cDNAs from fiber producing cells.

The final collection of clones were then subjected to northern analysis.For this analysis, poly(A)+ RNA from different tissues were denatured inthe presence of formaldehyde and size-fractionated on 1.5%agar/formaldehyde gels as described by John et al., supra. The RNAs werethen blotted to nitrocellulose and probed with ³² P-labelled inserts ofeach individual clone. The clones that showed hybridization to only RNAsfrom fiber cells were selected. This screen resulted in theidentification of cDNAs specific to five fiber specific genes. Allmanipulations on plasmid DNAs such as isolation, purification on cesiumchloride gradients, restriction digestion, insert purifications by gelelectrophoresis and electroelutions and ³² P-labelling bynick-translations have been described previously (Maniatis et al., supraand John et al., supra).

The cDNA library from the 10-day old cells was then screened using asubtractive hybridization procedure as follows. The ³² P-labelled cDNAfrom fiber was hybridized to excess biotinylated mRNA isolated from leaftissue. The cDNA-biotinylated mRNA hybrids and the excess biotinylatedmRNAs were separated from unhybridized cDNA by extraction with avidin inphenol:chloroform. The streptavidin was partitioned into the organicphase along with any biotinylated nucleic acid while the single-strandedcDNA remained in the aqueous phase. This procedure has been describedelsewhere, Duguid et al., Proc. Natl. Acad. Sci. USA, 85:5738-5742(1988).

Substractive hybridization screening of 4788 clones of the 10-day celllibrary with leaf cDNAs resulted in 800 clones not present in the leaf.These clones were then screened by cDNAs generated from ovule, flowerand root mRNAs. The results of this screening were 79 putativefiber-specific clones. The duplicate clones which hybridized to eachother were detected by the procedure of polymerase chain reaction(PCR)(Saiki et al., Science, 239:487-491 (1988)), Southern blotting andhybridization. The PCR reaction was carried out by first mixing 10microliters of bacterial culture of the cDNA clone added to 90microliters of distilled water. 20 microliters of that mixture was addedto a PCR reaction buffer of 50 mM KCl, 10 mM Tris-HCl pH 8.0, 2.5 mMMgCl₂, 0.01% gelatin, 200 μM each of dATP, dCTP, dTTP and dGTP, 12.5picomolar each of sense and antisense primers for pBR322, and 0.5 unitsof Taq polymerase. The final reaction volume was 52 microliters. The PCRreactions were carried out in a Perkin-Elmer-Cetus thermocycler.

The amplified DNA from the PCR reactions was separated by agarose gelelectrophoresis and blotted onto nitrocellulose by the method ofSouthern, J. Mol. Biol. 98:503-517 (1975). One or more bacterial clonesfrom the same group was amplified by the same procedure and the productsalso separated on agarose gel. The amplified insert DNAs were thenexcised from the gel and purified by electroelution. The purified DNAs,labelled with ³² p by nick-translation, were hybridized with theSouthern blot. Thus, the cross-hybridizing clones were identified inthis fashion. This procedure resulted in the identification of 19putative fiber-specific clones. The clones were further analyzed bynorthern blots. Three of the clones were found to be fiber-specific.Another five of the clones were found to be differentially expressed toa higher degree in fiber and to a lesser degree in other tissues.Fiber-specific cDNA clones were then used as probes to screen genomiclibraries and isolated cross-hybridizing genomic clones.

3. Characterization of Fiber-Specific cDNA Clones

In the following sections, we describe each of the fiber cDNAs, theircorresponding genes and their promoters. The nomenclature is as follows:CK=Coker, SI=Sea Island; FB15, FB10=15 or 10 day old bolls; the lastremaining letters and numbers signify the individual cDNA isolate.

a. CKFB15A1-E6 cDNA clone (E6 cDNA)

This cDNA clone for a fiber gene has an insert of 983 base pairs whichhybridizes to 1.0 and 1.1 Kb RNAs. The RNA is expressed in fiber and notin root. Flower leaf and ovule RNAs show weak hybridization.

The E6 RNA was found to be developmentally regulated. Its steady-stateconcentration increases immediately after anthesis. Our quantificationof E6 transcript in fiber using in vitro synthesized E6 RNA as acontrol, shows that 20 μg of RNA from 20-day-old fiber contains about3.5 ng of E6 RNA. Thus, E6 RNA is an abundant fiber RNA.

Hybrid selection translation experiments showed that E6 codes for twopolypeptides of 26 and 30 kD. The E6 clone cross-hybridizes with Pimaand Naked seed cotton fiber cell RNAs. The clone also cross-hybridizeswith a number of plants belonging to Gossypium species. Thus, DNAs fromPima and Sea Island (G. barbadense) PD3 and DP50 (G. hirsutum) andplants belonging to G.longicalyx and G. somalense all showedhybridization. In addition, plants belonging to another species offamily Malvaceae, the Hibiscus are also found to have conserved E6 gene.DNAs of H. sabdariffa L. cv., Rosselle, Kapok (Ceiba pentandra)belonging to family Bombacaceae, Hemp (Cannabis sativa) belonging tofamily Moraceae also showed hybridization to E6 gene. We also confirmedthat E6 or a homologous gene is present in Gossypium darwinii, Gossypiumherbaceum L. cv. Jayadhar and Tzuyung, Gossypium anomalum, G. australe,G. nelsonii, G. arboreim L., cv., Nanking and Liaochung, G. thurberi, G.davidsonii, G. raimondii, G. stocksii, G. somalense, G. longicalyx, andF. bickii. Thus, the E6 sequence is conserved in most of the plantsbelonging to family Malvaceae and also found in two other familiesBombacaceae and Moraceae. Many of these plants produce seed hair or bastfiber. Interestingly, we did not detect E6 hybridization in the DNAs ofsoybean, corn, tobacco or the cellulose producing bacterium Acetobacter(A. xylinum). These studies imply that E6 gene may have functions in theformation of seed hair or bast fiber cells.

The complete nucleotide sequence of E6 insert is presented as SEQ ID NO:2. This sequence contains a long open reading frame extending fromposition 1 to position 748. On this same open reading frame, startcodons appear at positions 34, 61 and 94. If the first codon is theinitiation site for the protein, the 714 nucleotide reading frame wouldyield a 238 amino acid protein. E6 cDNA clone was deposited with ATCC atAccession Number 67809.

SEQ ID NO: 2 also contains an additional 84 residues and a stretch ofpoly(A) that originate from clone PCKFB15-B3. This clone is identical topCKFB15A1-E6 except for the presence of additional residues at the 3'end.

b. CKFB15A1-H6 cDNA clone (H6 cDNA)

H6 cDNA hybridizes to a developmentally regulated RNA of 950 bases. H6RNA was not detected in leaf, flower, ovule and root. The H6 clonecross-hybridizes to Pima, PD3 and Sea Island DNAs and is encoded by oneor two genes in the cotton genome. The H6 clone had an insert of about500 base pairs.

To obtain a full length cDNA clone, primer extension of H6 clone mRNAwas conducted using an oligomer and fiber cell mRNA by the protocoldescribed by Dean et al., Nucleic Acid Res. 15:4655-4668 (1987). Theprimer-extended product was then cloned into the Pst 1 site of dG-tailedpBR322. The complete sequence of H6 insert clone and the primer extendedH6 (CKFBH6-10) were determined. Together, these two sequences make upthe complete 913 base pair sequence of H6. This sequence, SEQ ID NO: 3below, has a single long open reading frame with an initiation codon atposition 71. Sequence analysis using a Genetics Computer Group softwarepackage (University of Wisconsin) for identifying protein codingsequences also suggest a single protein coding region between residuepositions 71 and 710. The nucleotide-derived amino acid compositionshows a proline rich peptide (35 mole % proline) of 214 amino acids. Atotal of five amino acids (alanine, proline, leucine, serine and valine)make up 74.3 % of the protein. The sequence includes 17 pentapeptiderepeats of X-Y-Pro-Pro-Pro repeat units where X and Y are serine,alanine or theronine. The H6 protein is clearly distinct from previouslyknown proteins of plant cell walls, such as extensin. H6 cDNA clone wasdeposited with ATCC at Accession Number 67810.

c. CKFB15A1.C12 cDNA clone (C12 cDNA)

C12 RNA is 1.1 Kb bases long and is developmentally regulated. It is notexpressed in root, leaf, ovule and flower. The C12 clone crosshybridizes with Pima, PD3 and Sea Island genomic DNAs, and is encoded byone or two genes. The C12 clone has an insert of about 659 base pairs.The sequence is presented as SEQ ID NO: 4 below. The C12 cDNA clone hasbeen deposited with ATCC at Accession Number 67808.

d. CKFB15A1-B8 cDNA clone (B8 cDNA)

B8 RNA is 1100 bases long and is developmentally regulated. It is notexpressed in leaf, root, ovule and flower. B8 cross-hybridizes to Pima,PD3 and Sea Island genomic DNAs and is encoded by one or two genes. TheB8 cDNA clone has an insert of 690 bp, the sequence of which ispresented at SEQ ID NO: 5 below. It has been deposited with ATCC atAccession Number 67807.

e. CKFB1O-B12 cDNA clone (B12 cDNA)

B12 cDNA hybridizes only to fiber RNA in northern analysis. Thetranscript size is 1 Kb. The 727 base pair insert in B12 has beensequenced and is presented as SEQ ID NO: 6 below. The developmentalpattern of expression of the clone showed that maximum concentration ofB12 mRNA is present 10 to 20 days after anthesis. The concentration ofB12 RNA in 24 day old cotton fiber cells is very low.

f. CKFB1O-A11 cDNA clone (FB10-A-11 cDNA)

FB10-A-11 RNA is also fiber-specific. The cDNA insert size is 1 Kb. TwomRNA transcripts (1.1 and 0.9 Kb) from fiber cells hybridize toFB10-A-11. The sequence for FB10-A11 is presented as SEQ ID NO: 7 below.

g. CKFB1O-D7 cDNA clone (D7 cDNA)

This cDNA clone hybridizes to an RNA of about 500 bases in length. It isnot detected in ovule, leaf, flower or root RNA from cotton. Thesequence of this cDNA clone is presented as SEQ ID NO: 8 below.

h. CKFB1O-C2 cDNA clone (C2 cDNA)

This cDNA clone hybridizes to an mRNA highly expressed in cotton fibercells but also detected as weakly present in petal tissues. The cDNAinsert is 668 bp and hybridizes to an RNA of 1.1 Kb. The sequence islisted as SEQ ID NO: 9.

i. CKFB1O-C12 cDNA clone (C12 cDNA)

The cDNA clone C12 has an insert size of 609 base pairs. The transcriptis expressed in fiber cells, but is also expressed at low levels inpetal and pollen. The cDNA hybridizes to an mRNA of 1.1 Kb. The sequenceof CKFB1O-C12 is shown in SEQ ID NO: 10 below.

j. CKFB1O-C1 cDNA clone (C1 cDNA).

This clone hybridizes to a transcript of 450 base pairs in fiber cells.The cDNA also hybridizes very weakly to transcripts in petal and pollen.The insert size is 432 base pairs. The sequence is presented at SEQ IDNO: 11 below.

k. CKFB1O-A8 CDNA clone (A8 cDNA)

This cDNA clone has an insert size of 320 base pairs and hybridizes to a1 Kb mRNA in fiber cells. The clone also exhibits weak hybridization toleaf and to petal RNA. The sequence of the insert is presented in SEQ IDNO: 12.

l. CKFB1O-A9 cDNA clone (A9 cDNA)

The cDNA clone A9 has an insert of 399 base pairs and hybridizes to anRNA of 750 bases in fiber cells. The clone exhibits weaker hybridizationto RNAs from other tissues. The sequence is presented at SEQ ID NO: 13below.

m. CKFB1O-D4 cDNA clone (D4 cDNA)

Clone D4 hybridizes strongly to 10 day fiber RNA and very weakly topetal RNA. Its transcript size is 500 bases and has an insert size of455 bp. This sequence is presented at SEQ ID NO: 14 below.

n. CKFB1O-B6 cDNA clone (B6 cDNA)

B6 cDNA hybridizes to RNA of fiber. It also shows weak hybridization toleaf RNA. It has an insert size of 1.1 Kb and transcript size of 1.2 Kb.The sequence of B6 cDNA is presented at SEQ ID NO: 15.

o. CKFB1O-A12 cDNA clone (A12 cDNA)

A12 cDNA hybridizes to fiber RNA only. It has an insert size of 868 bpand hybridizes to a RNA of 900 bases. The sequence of A12 cDNA ispresented at SEQ ID NO: 16.

p. CKFB15-E9 cDNA clone (E9 cDNA).

This clone hybridizes strongly to fiber RNA and weakly to petal RNA. Ithas an insert size of 1283 bp. The sequence is presented at SEQ ID NO:17 below.

q. SIFB-H8 cDNA clone (H8 cDNA)

H8 cDNA hybridized to fiber RNA, but not to cotton leaf, root or ovule.The H8 sequence is presented at SEQ ID NO: 18 and contains an 878 bpinsert.

r. SIFB-H4 cDNA clone (H4 cDNA)

mRNA hybridizing to H4 cDNA is expressed in fiber and to a much lesserextent in petals. The sequence of H4 cDNA is presented at SEQ ID NO: 19.

As will become apparent from the following, these cDNA clones can beused to obtain genomic clones containing fiber-specific promoters. Planttransformation vectors can therefore be constructed to transfer genesconnected to these promoters into fiber-producing plants and express thegene in a fiber-specific manner.

Some of the RNA species represented by the above-identified cDNA clonesare partially expressed in other tissues, such as petal or leaf. If apromoter is desired that directs expression in fiber and another planttissue, one of these cDNA clones might be preferentially selected tosearch for this promoter. For example, E9 and H4 cDNAs hybridize weaklywith petal RNA. If one wished to obtain a promoter that preferentiallypromotes expression of genes in fiber, but also promotes expression to alesser extent in petal, one would use the E9 sequence or the H4 sequenceas probes.

4. Preparation of Genomic DNA and Genomic Clones

Genomic DNAs from Sea Island cotton, Coker 312 cotton and Kapok wereprepared according to the methods described in Current Protocols inMolecular Biology, (Eds. Ausbel, F. M. et al.) Wiley, (1987), pp.2.3.1-2.3.3, with the following modification: The frozen plant materialwas homogenized in extraction buffer containing 1% polyvinylpyrrolidone. The purified genomic DNA was digested with restrictionendonucleases and transferred to nitrocellulose filters by the Southernblotting technique. Southern, (supra).

The filters were then probed with nick-translated inserts of thefiber-specific cDNA clones previously identified. The hybridization andblot washing conditions are described in John et al. (supra). TheSouthern hybridization results showed that each of the cDNA cloneshybridized to only a few (one or two) bands in the genomic DNA. Thisresult indicates that there are only one or two genes corresponding tothese cDNAs in the cotton genome.

Sea Island cotton, Coker 312 cotton and Kapok genomic libraries wereprepared by Clonetec, Inc., of Calif., in EMBL-3 or lambda DASH vectors.For the preparation of the library, the DNA was partially digested withMbo I. 8-22 Kb DNA fragments were cloned into the Bam HI site of thevector. Inserts 10-15 Kb were present in the phages. The inserts wereexcised by either Sal I for EMBL-3 and lambda DASH or Eco R1 for lambdaDASH. The genomic libraries were plated on E. coli NM 538 as describedin Current Protocols in Molecular Biology, (supra.) The genomic librarywas screened with ³² P-labelled inserts of the fiber-specific cDNAclones after transferring the library to nitrocellulose filtersaccording to the methods described in Current Protocols, (supra) andJohn et al., (supra). By this method, genomic clones containingsequences homologous to the fiber-specific cDNA clones were isolated.

Subcloning of these genomic DNA inserts into plasmid or phagemid vectorswas done using standard protocols. Ligated DNAs were transformed intoEscherichia coli strain XL-1 Blue (Stratagene). Recombinant clones wereselected on the basis of blue/white selection on X-gal, IPTG(5-bromo-4-chloro-3-indoyl-beta-D-galactophyranoside;ispropyl-beta-thio-galactophyranoside) plates. The plasmid sizes of therecombinant clones were then analyzed by SDS-agarose gel electrophoresis(Sekar, V., Biotechniques, 5:11-13, (1987)). The inserts of the cloneswere further characterized by restriction mapping and Southern analysis.Current Protocols in Molecular Biology, (supra). If necessary, furthersubcloning of smaller restriction fragments that contain the cDNAhybridizing regions was also undertaken. The above protocols enable oneto determine the approximate boundaries of a given gene.

The nucleotide sequence of the gene and the corresponding cDNAs wereanalyzed by computer programs to determine, among other things, thetentative coding region, presence of introns and exons, 5' and 3'non-coding regions and putative promoter regions. The software that wehave used for this purpose is that of Genetics Computer Group (GCG),Madison. Once the detailed sequence analysis was performed and variousputative structural components of the gene were identified, we were ableto confirm these findings by various experiments. For example, we used achimeric marker gene construct that includes the promoter fragment totransform a test cell. By observing the presence or absence of themarker gene activity, one can analyze the promoter function of that DNAfragment.

5. Characterization of Genomic Clones

Once the genomic clones were identified, they were subcloned intoplasmid or phagemid vectors for easy manipulation, as discussed above. Aschematic representation of the promoter fragments that we characterizedis given in FIG. 1.

The nomenclature is as follows: EMBL and DASH=Lambda vector; SI=SeaIsland, CK=Coker; CP=Ceiba pentandra; next to last two characters=thecDNA clone that hybridized to the genomic clone; the last numbers orcharacters correspond to different genomic clones from a given library.The following fragment sizes are approximate.

The genomic clones we obtained and assayed are described below inTable 1. When the fragments were initially cloned, a Sal I site wasadded to the fragment by the cloning vector. The designation "Sal I (MboI)" in the following table is to emphasize that a naturally occurringMbo I site exists adjacent to the artificial Sal I site. (The genomicfragments were originally created by a partial Mbo I digest.) All otherrestriction sites in Table 1 and in FIG. 1 are naturally occurringgenomic sites.

                                      TABLE 1                                     __________________________________________________________________________                        FRAGMENT ASSAYED                                          GENOMIC CLONE                                                                            SUB CLONE                                                                              FOR PROMOTER SEQUENCE ID NO:                              (Insert Size)                                                                            (Insert Size)                                                                          ACTIVITY     AND DESCRIPTION                              __________________________________________________________________________    EMBL-SI-E6-2A                                                                            SKSIE6-2AH3                                                                            Sal I (Mbo I)/Nco I                                                                        SEQ ID NO: 20 contains 541 bp of                                              promoter, 33 bp of 5'                        14.2 Kb Sal I                                                                            6.4 Kb   4.5 Kb       noncoding, 741 bp of coding and 332 bp                                        of 3' noncoding                              (Mbo I)    Sal I/Hind III                                                     EMBL-SI-E6-3B                                                                            SKSIE6-3B                                                                              Sal I (Mbo I)/Nco I                                                                        SEQ ID NO: 21 contains 581 bp of                                              promoter, 33 bp of 5'                        15.0 Kb Sal I                                                                            5.1 Kb Sal I                                                                           2.7 Kb       noncoding region, 741 bp of coding                                            region, and 313 bp of                        (Mbo I)                          3' noncoding region.                         DASH-CK-E6-1A                                                                            SKCKE6-1A                                                                              4.1 Kb       SEQ ID NO: 22 contains 567 bp of                                              promoter, 33 bp of 5'                        12 Kb Sal I                                                                              12 Kb Sal I                                                                            Nco I        noncoding region, 717 bp of coding                                            region, and 301 bp of                        (Mbo I)                          3' noncoding region.                         DASH-CK-E6-4A                                                                            SKCKE6-4A                                                                              3.9 Kb       SEQ ID NO: 23 contains 512 bp of                                              promoter, 37 bp of 5'                        12.2 Kb Sal I                                                                            8.0 Kb Sal I                                                                           Sal I (Mbo I)/Nco I                                                                        noncoding region, 726 bp of coding                                            region, and 303 bp of                        (Mbo I)                          3' noncoding region.                         EMBL-CP-E6-3A                                                                            SKCPE6-3A-RV                                                                           3.2 Kb Sal I (Mbo I)/                                                                      SEQ ID NO: 24 contains 421 bp of                                              promoter including the 5'                    15.3 Kb Sal I                                                                            4.8 Kb   Nco I        untranslated mRNA leader, 873 bp of                                           coding region and 324                        (Mbo I)    Sal I/EcoRV           bp of 3' untranslated region.                EMBL-SI-H6-4                                                                             SKSIE6-H6-RI                                                                           321 bp Fsp I/Sal I                                                                         SEQ ID NO: 25 contains 250 bp of                                              promoter, 71 bp 5'                           13 Kb Sal I                                                                              1.9 Kb Sal I/                                                                          (Mbo I)      untranslated mRNA leader, 1226 bp of                                          coding region                                (Mbo I)    Eco RI                including a 583 bp intron and 437 bp of                                       3' untranslated region.                      EMB2-SI-B12                                                                              SKSI-B12-H3                                                                            3.1 Kb Eco RI/Sty I                                                                        SEQ id NO: 26 contains 699 bp of                                              promoter, 139 bp of 5'                       15 Kb Sal I                                                                              7.3 Kb Hind III       untranslated mRNA leader, 841 bp of                                           coding region which                          (Mbo I)                          includes two introns (76 bp and 82 bp                                         each) and 735 bp of 3'                                                        untranslated region.                         EMBL-SI-All-B                                                                            SKSIA11-B                                                                              2.1 Kb Sal I (Mbo I)/                                                                      SEQ ID NO: 27 contains 279 bp of                                              promoter                                     17.0 Kb Sal I                                                                            4.9 Kb Sal I                                                                           Eco RI                                                    (Mbo I)                                                                       EMBL-SI-B8 SKSIB8-HI                                                                              2.2 Kb       SEQ ID NO: 28 includes 287 bp of                                              promoter including mRNA                      19.0 Kb Sal I                                                                            9.5 Kb Sal I/                                                                          Bam HI/BstBI 5' leader, 1473 bp of coding region                                           (which includes a 510                        (Mbo I)    Bam HI                bp intron), and a 758 bp 3' region.          EMBL-SI-B6-1A                                                                            SKSSIB6-1A                                                                             2.3 Kb       SEQ ID NO: 29 contains 562 bp of                                              promoter.                                    of 12.3 Kb Sal I                                                                         12.3 Kb Sal I                                                                          Bam HI/Xho I                                              (Mbo I)                                                                       EMBL-SI-E9 SKSI-E9  7.0 Kb Xho I/                                             12.4 Kb Sal I                                                                            12.4 Kb Sal I                                                                          Nco I                                                     (Mbo I)                                                                       EMBL-SI-A12-A                                                                            SKSIA12-A                                                                              3.5 Kb Sal I (Mbo I)/                                     11.5 Kb Sal I                                                                            6.7 Kb Sal I                                                                           Sma I                                                     (Mbo I)                                                                       __________________________________________________________________________

6. Characterization of Promoter Activity (In General)

Once the DNA was purified from the phage genomic clones (Ausubel et al.,pp. 1.10.1 to 1.13.6) the insert DNAs (10 to 15 Kb) were characterizedin terms of their restriction maps (supra, pp. 3.1.1 to 3.3.2). Thedifferent restriction fragments were separated on agarose gels andSouthern blotted. The blots were then be hybridized to cDNA probes. Thisprocedure enabled us to identify smaller fragments (about 1 to 10 Kb)that contained the homologous cDNA sequence. The fragment was thensubcloned (supra, pp. 3.16.1 to 3.16.11) into plasmid vectors such aspGEM5zf (Promega, Madison) or Bluescript SK, KS (Stratagene, Calif.).All further manipulations, such as promoter identifications,transcription maps and gene size determinations were done using thesubclones.

Mapping the gene transcripts by nuclease protection may also be done.Single-stranded DNA probes may be generated from the Bluescriptsubclones and hybridized to poly(A)+ RNA from fiber cells. Thehybridized portions that are protected from nuclease action will bedetermined as described by Calzone et al. in Methods in Enzymology, Vol.152 (Eds. Berger, S. L. and Kimmel, A. R.), 1987, pp. 611-632.Furthermore, mapping the 5' termini by cDNA primer extension is alsodescribed (supra, pp. 629-632). These strategies will determine the sizeof the gene, as well as precise boundaries of the gene transcript, orcoding region for the fiber gene, in the subclone. Portions of the DNAmay then be sequenced if desired.

It was necessary to determine whether the isolated DNA fragments hadpromoter activity. The genomic fragments we assayed are described inFIG. 1.

A GUS fusion construct was used to identify the cotton promoters. First,the transcription start site of the mRNA was determined by primerextension method, as described by Calzone et al. (supra). The subclonedgene fragment and a short restriction fragment or an oligomer at the 5'end was used in the primer extension. A beta-glucuronidase (GUS) codingsequence along with necessary termination signals, as well a 5' leadersequences, was used with an upstream 2 -3 Kb DNA fragment from thetranscription or translation start site. The GUS sequence is alreadyreadily publicly available (ATCC 67641).

It has been demonstrated that a GUS gene construct with a cauliflowermosaic virus 35 promoter (CaMV35s) promoter is functional in plantcells. Thus, when this construct was introduced through particlebombardment (described by McCabe, D. et al., Bio/Technology, 6:923-926,1988) into a plant, GUS activity was observed. If the constructcontaining an unknown DNA is found to be active in expressing GUS, thenit can be concluded that the DNA fragment contains a promoter thatdirects the expression of GUS gene. Using the GUS assay, we determinedthat the sequences identified in Table 1 had promoter activity. FIG. 2describes the various levels of promoter activity we found.

Using the Bluescript subclone and exonuclease/mung bean deletionprocedure in which a series of clones with differing lengths of the 5'fragment are generated, one can identify minimum lengths of 5' DNAnecessary to express the gene in fiber cells. These types of procedureswill enable one to identify promoters from all genomic clones. Based onthis knowledge, one can construct various developmentally regulatedexpression vectors containing fiber genes of interest and introduce theminto plants.

7. Determination of Promoter Activity

A chimeric gene construct was made using the putative promoter and thereporter gene beta-glucuronidase (GUS) of E. coli. GUS catalyzes thecleavage of 5-bromo-4-chloro-3-indoyl glucuronide (X-Gluc). The indolederivative produced by this cleavage undergoes oxidative dimerization toform a blue dye. Cells that produce this blue dye can be detectedeasily. The GUS marker system has been described in detail by Jefferson,et al., in Proc. Natl. Acad. Sci. USA 83:8447-8451 (1986) and in PlantMol. Biol. Rep. 5:387-405 (1987). The GUS gene is publicly available(ATCC Accession No. 67641).

Chimeric plasmids were constructed by litigating a promoter-less GUScoding region along with a transcription termination signal Nos(A) atthe 3' end into a vector cassette as a Nco I/Sal I fragment. An AMV 5'untranslated leader is added to the 5' end of the GUS gene as a NcoI/Xho I fragment. This construct (p2117, FIG. 3) contains unique Xho Iand Nco I sites for introducing putative promoters. For example, if acauliflower mosaic virus 35s (CaMV 35s) promoter is ligated into Xho Ior Nco I site and the resulting plasmid (p2119) is introduced into plantcells by particle acceleration, GUS expression can be detected. Ellis etal., Plant. Mol. Biol. 17:19-27, 1991. We have tested GUS expression byhistochemical staining as well as quantitative measurements (Jefferson,R. A., supra).

The assay involves transforming a test cotton tissue, such as hypocotyl,with different plasmids. We transformed the hypocotyl tissue via aparticle-mediated transformation method disclosed in U.S. Pat. No.5,015,580, hereby incorporated by reference.

We analyzed our reporter gene constructs in two ways, throughhistochemical staining and through fluorogenic analysis. Thehistochemical staining gave a quick "yes or no" answer while thefluorogenic analysis provided quantitative data.

(a) Histochemical Staining

Histochemical localization of beta-glucuronidase activity in planttissues is achieved by incubating freshly cut, transformed tissuesections in a solution containing 5-bromo-4-chloro-3-indoyl glucuronide(X-Gluc). X-Gluc is prepared by dissolving 5 mg of X-Gluc in 50 μl ofdimethyl formamide and diluting it to 10 ml with 50 mM sodium phosphatebuffer pH 7.0. After staining (1-3 hours at 37° C.), the tissue sectionswere rinsed off with 70% ethanol. Cells containing an active GUS geneturn blue.

(b) Fluorogenic Assay

The quantitative assay for GUS activity depends on the cleavage of4-methyl umbelliferyl glucuronide (MUG) by the GUS enzyme into afluorogenic product 4-methyl umbelliferone (MU). MU is fluorescent whenits hydroxyl group is ionized. (Jefferson, R. A. supra). The fluorogenicassay is carried out as follows. Plant tissue was homogenized inextraction buffer (50 mM NaH₂ PO₄, pH 7.0, 10 mM EDTA 0.1% Triton X-100,0.1% sodium lauroyl sarcosine, 10 mM β-mercaptoethanol). We includedproteinase inhibitor PMSF at a final concentration of 20 μg/ml and 1%insoluble PVP. The extract, after centrifugation (300 μl) was added to 1ml of MUG buffer. The MUG buffer is made up of 1 mM MUG in the aboveextraction buffer. The mixture is incubated at 37° C. and at time points0, 20, 40, and 60 min. an aliquote (100 μl) is withdrawn and added to 1ml stop solution (0.2 M Na₂ CO₃). The fluorescence at each time point ismeasured in a fluorocalorimeter (excitation at 365 nm, emission at 455nm) Protein concentration of the plant extract is determined by Bradfordassay using a test kit from Bio-Rad Laboratories (M. Bradford, Anal.Biochem. 72:248-254, 1976). The fluorimeter is calibrated with freshlyprepared MU standards. The results are given as pmole MU/mg/min. FIG. 2describes the various results with our twelve promoter fragments.

The above system for the detection of promoters, namely transientexpression of chimeric GUS plasmids introduced into hypocotyl tissuesthrough particle bombardment, appears to be limited in that no tissuespecificity of expression is observed for any of the promoters tested.Thus, promoters (LAT 52, Cab) that were proven to be tissue specific inheterologous stable transgenic plants (Twell, et al., Mol. Gen. Genet.217:240-245 (1989); Ha and An, Proc. Natl. Acad Sci. USA 85:8017-8021(19.88)) are found to express GUS transiently when introduced intocotton hypocotyls by particle bombardment.

The following control experiments were conducted along with each of thepromoter assays.

1. Plasmid p2117 was introduced into cotton or soybean hypocotyl tissuesas a negative control. Because p2117 contains a promoter-less GUS gene,no GUS activity should be detected. FIG. 3 describes p2117.

2. The tissue was bombarded with p2119. p2119 contains a GUS gene drivenby 35s promoter. This is a positive control, for GUS should always beexpressed.

3. The DNA fragment being tested for activity is introduced intohypocotyl tissue. This is to demonstrate that the fragment in questionhas no GUS-like activity.

Using the above procedures and protocols, we have demonstrated promoteractivity in the twelve promoter fragments listed in FIG. 1. Thepromoters exhibited different levels of expression, as FIG. 2 indicates.If one wished to create a transgenic plant with high or low expressionof a certain transgene, FIG. 2 would allow one to select the appropriatepromoter.

8. Transformation of Plants

The most common methodology used for the transformation of cells ofdicot plant species involves the use of the plant pathogen Agrobacteriumtumefaciens. A. tumefaciens harbors a plasmid, referred to as thetumor-inducing or Ti plasmid, which has the natural ability to transfera segment of itself, referred to as the T-DNA (transfer-DNA), into thegenome of infected plant cells. Wild-type A. tumefaciens use thisability to genetically transform infected cells of plants so that theplant cells become tumorous, and synthesize one of a series ofcompounds, known as opines, which can be metabolized by the infecting A.tumefaciens. Several investigators have found that by removing the bulkof the T-DNA from the Ti plasmid harbored by A. tumefaciens, and byreplacing that T-DNA with a foreign gene construction, the Agrobacteriumcan transform infected plant cells with the foreign gene in such afashion that the resultant cells are not tumorous, as plant cellsinfected with wild-type A. tumefaciens normally are. The foreign geneconstruction is then included in the cells of a whole plant regeneratedfrom the transformed cells and is then inherited in a simple Mendelianmanner. The construction can thus be treated as any inheritable traitfor crop breeding purposes. The transformation and regeneration ofcotton plants by Agrobacterium transformation has been achieved andreported. Umbeck et al. (supra) and Firoozabady et al. (supra).

Other methods of plant transformation, such as transformation byaccelerated particle carried DNA, are now available (McCabe et al.supra). An apparatus capable of performing particle-mediatedtransformation is commercially available from BioRad. In any event, oncethe creation and assembly of plant expression vectors including fiberspecific gene sequences is accomplished, the transformation andregeneration of cotton plants with these expression vectors is withinthe ability of one of ordinary skill in plant genetic engineering, andis not dependent on the method of transformation.

9. Characterization of Cotton Genes

As a specific Example of our characterization of fiber-specificpromoters, our characterization of the E6 genes is disclosed in detail.Brief descriptions of other genes and promoters follow.

Two E6 genes from cultivar Sea Island (G. barbadense) and two genes fromCoker 312 (G. hirsutum) were isolated and examined. These four geneshave homology to each other. Detailed structural characterization ofthese genes is presented below.

a. Sea Island E6-2A Gene and Promoter.

The phage EMBLSIE6-2 contains a 14.2 Kb insert. Restriction mappingfollowed by Southern analysis showed that a 9.5 Kb Sal I fragmenthybridized to E6 cDNA. The 9.5 Kb Sal I fragment was subcloned into Sk⁺vector (pSkSIE6-2A). Further subcloning resulted in a shorter clone (6.4Kb) pSKSIE6-2AH3. About 541 bp of promoter region, 33 bp of 5' noncodingregion, 741 bp of coding region, and 332 bp of 3' noncoding region havebeen sequenced and is described in SEQ ID NO: 20. In order to identifythe promoter region, plasmid pSKSIE6-2AH3 was digested with Nco I andBam HI to liberate three fragments. The vector and upstream 5' noncodingfragment of the gene (4.5 Kb insert and 3 Kb vector) was gel purifiedand subjected to Elutip-d column elution.

Plasmid 2117 is a promoter-less GUS marker gene cassette. A circular mapof the plasmid is shown in FIG. 3. When a 35s promoter is added top2117, the marker gene is able to express GUS in plant tissue. Plasmid2117 was digested with Nco I and Bam HI and gel purified. The 2 Kb NcoI/Bam HI fragment containing the GUS gene was ligated to the 7.5 Kb E6genomic fragment and transformed into XL-1 Blue cells. Recombinantclones (p2117-E6) were identified by SDS-agarose gels. However, thisconstruct lacks a poly(A)-addition signal. Therefore, a 277 bp Bam HIfragment of p2117, which contains the poly(A)-addition signal, wasligated into the Bam HI site of p2117-E6. The orientation of the poly(A)addition signal in p2117E6P-2A was then determined by restrictiondigestion analysis and the chimeric plasmid containing the correctorientation was selected. The construction of the plasmid is shown inFIG. 4.

Plasmid p2117E6P-2A was then introduced into cotton hypocotyls throughparticle acceleration method (McCabe, D., et al., Bio/Technology,6:923-926 (1988)). As mentioned above, control experiments used plasmidsp2119, p2117, and pSKSIE6-2AH3. Plasmid p2119, a GUS construct with the35s promoter gave positive GUS activity as expected. Plasmid 2117 (noplant promoter) and plasmid SKSIE6-2AH3 (no GUS gene) gave no GUSactivity. Plasmid p2117E6P-2A which contains the region 5' to the E6gene and the GUS gene, showed GUS activity. This result suggests that anactive promoter element is located in the 4.5 Kb Nco I/Sal I DNAfragment.

b. Sea Island E6-3B Gene and Promoter.

A second phage (insert 15 Kb) hybridizing to E6 cDNA contains a Sal Ifragment (5.1 Kb). This fragment was subcloned into SK⁺ vector. Thisplasmid (pSKSIE6-3B) was characterized in terms of the region to whichE6 cDNA hybridize. 581 bp of promoter region, 33 bp of 5' noncodingregion, 741 bp of coding region, and 313 bp of 3' noncoding region areshown in SEQ ID NO: 21. Promoter fragment identification of pEKSIE6-3Bgene was carried out as follows. After determining the coding region andthe putative promoter region in pSKSIE6-3B it was digested with Nco Iand Xba I and the resulting plasmid (approximately 5.7 Kb) gel purified.Similarly the promoter-less GUS gene cassette was digested with NCOI/Xba I and gel purified. The GUS gene was then ligated to plasmidpSKSIE6-3B. Construction of p2117 E6P-3B is shown in FIG. 5. Asdescribed for p2117 E6P-2A, various control plasmids along withp2117E6P-3B were introduced into hypocotyl to test promoter activity.p2117E6P-3B showed GUS expression indicating that the 2.7 Kb Nco I/Sal IE6-3B fragment contains the promoter of E6-3B gene. Relative expressionlevel of E6-3B promoter is shown in FIG. 2.

c. Coker E6 Gene pSKCKE6-1A and Promoter.

The screening of a Coker 312 genomic library with pCKFb15A1-E6 cDNAresulted in the identification of two phages, DASHCKE6-1A andDASHCKE6-4A. Inserts from these two phages were subcloned into SK⁺vector.

A 12 Kb insert from phage DASHCKE6-1A was subcloned into SK⁺ vector togive pSKCKE6-1A plasmid. A detailed restriction map of the insert wasprepared and its comparison with the map of pCKFb15A1-E6 enabled us tolocate the coding region of Coker E6 gene in pSKCKE6-1A. About 567 bp ofpromoter, 33 bp of 5' noncoding region, 717 bp of coding region, and 301bp of 3' noncoding region are included in the sequence and is shown inSEQ ID NO: 22. A 4.1 Kb Nco I fragment upstream of the coding regionmost likely contains the promoter of this gene. To confirm this, the NcoI fragment was ligated into the Nco I site of p2117 and the constructwas tested for GUS activity as described earlier. The resultsdemonstrated that the 4.1 Kb fragment contains a promoter.

d. Coker E6 Gene pSKCKE6-4A and Promoter.

An 8.0 Kb Sal I fragment from a second phage (12.2 Kb insert) hybridizedto E6 cDNA. This fragment was subcloned into SK⁺ vector (pSKCKE6-4A).512 bp of promoter region, 37 bp of 5' noncoding region, 726 bp ofcoding region, and 303 bp of 3' noncoding region have been sequenced andis shown in SEQ ID NO: 23. In a manner similar to our work withpSKCKE6-1A, we determined the location of the promoter of this gene andsubcloned a 3.9 Kb Nco I/Sal I fragment into p2117 at the Xho I/Nco Isites. The resulting plasmid was then tested for GUS activity and wasfound to be able to direct synthesis of GUS enzyme in plant tissue.

e. Characterization of Cebia pentandra (Kapok E6 Gene and Promoter.

An EMBL-3 genomic library of Cebia pentandra (Kapok) was screened withE6 cDNA. Four hybridizing phages were identified. One of the phageinserts was subcloned into Sal I site of Bluescript vector SK⁺. Theresulting clone, pSKCPE6-3A (15.3 Kb insert) was characterized byrestriction analysis and Southern blotting. We identified a Sal I - EcoRV fragment (4.8 Kb) that hybridizes to CKFB15A1-E6. This fragment wassubcloned into the Sal I - Eco RV site of Bluescript vector resulting inclone pSKCPE6-3A-RV. A 1.6 Kb stretch of DNA was sequenced and is shownin SEQ ID NO: 24. Comparison of cotton E6 cDNA sequence and Kapok E6gene sequence revel 84.3% homology.

Plasmid SKCPE6-3A-RV was digested with Nco I and Bam HI and thefragments separated on agarose gels. The larger of the DNA fragments(6.2 Kb, a 3.2 Kb insert and a 3 Kb vector) was electroeluted andpurified. Similarly, GUS gene containing p2117 was digested with Nco Iand Bam HI and the 1.8 Kb GUS fragment isolated and purified. The twoDNA fragments were then ligated and transformed into X1-1 Blue cells.Recombinant clones were identified by DNA analysis on SDS-agarose gels(Sekar, et al., supra). A Nos poly(A) signal region from p2117 wasisolated by Bam HI digestion. This fragment was ligated into the Bam HIsite of the above Kapok E6-GUS plasmid after linearization with Bam HI.The resulting clones was analyzed for correct orientation of the Nospoly(A) addition signal by restriction digestion analysis. A circularmap of pCPE6-2117 is shown in FIG. 6.

Plasmid CPE6-2117 was introduced into cotton hypocotyls through particleacceleration method (McCabe et al., (supra). In duplicate experiments,pSKCPE6-RV, p2117 and p2119 were introduced into hypocotyls. Asdescribed earlier, GUS activity was assayed. As expected, pSKCPE6-RV andp2117 showed no GUS activity while p2119 gave positive results. Theplasmid CPE6-2117 showed GUS activity, confirming that the 3.2 Kb SalI/Nco I fragment contains the promoter for CPE6 gene.

f. Characterization of Sea Island H6 Gene and Its Promoter

A 13 Kb Sal I fragment from the H6 genomic clone, EMBLSIH6-4, wassubcloned into Bluescript SK⁺ vector and designated pSKSIH6-4. Afteridentifying the cDNA hybridizing region in pSKSIH6-4, the H6 gene wasfurther subcloned by digesting the plasmid with Eco RI, purifying thefragments, and ligating them to SK⁺ vector. One of the resultingplasmids, designated pSIH6-4R1, contains the H6 gene. The completesequence of the H6 coding region as well as the 300 bases upstream fromthe initiation codon were determined. These sequences are presented atSEQ ID NO: 25. Digestion with the enzyme Fsp I releases this 300 bpregion, together with 250 bp of vector sequence. The 550 bp fragment wastested for promoter function in a similar fashion described for E6genes, and is found to be active. FIG. 7 is a diagram of the H6 cDNA.FIG. 8 is a diagram of the H6 gene. FIG. 9 is a diagram of plasmidpH6-2117.

g. Characterization of B12 Gene and Its Promoter

A genomic clone corresponding to pCKFB10-B12 was isolated from the SeaIsland genomic library. The 15 Kb insert from the phage, EMBLSI-B12, wassubcloned into an SK⁺ vector as a Sal I fragment. Further subcloning ofthe insert as a Hind III fragment (7.3 Kb) into SK⁺ vector resulted inplasmid pSKSI-B12H3. The sequence of B12 gene and 5' flanking promoterregion is shown in SEQ ID NO: 26.

The promoter fragment of the gene was identified as follows. Thefragment corresponding to the 5' end of the cDNA in the B12 gene wasexcised as an EcoR1/Sty I fragment (3.1Kb) and gel purified. After T4DNA polymerase treatment, Nco I linkers were added and then ligated tothe Nco I site of p2117. Orientations of the fragment in relation to GUSgene was determined by restriction mapping and clones with the correctorientation was selected. pB12-2117 was introduced into hypocotyl tissuethrough particle bombardment for GUS activity measurements.

h. Characterization of Sea Island A-11 Gene and Its Promoter

The fiber cDNA A-11 was used to screen a Sea Island genomic library andisolate phage EMBLSI-A11A and EMBLSI-A11B. The insert of the phageEMBLSI-A11B (17 Kb) was subcloned into SK⁺ vector and subjected torestriction mapping and Southern analysis. A 4.9 Kb Sal I fragment thathybridized to pCKFB10-A11 cDNA was then subcloned into SK⁺ vector(pSKSIA11-B) and the cDNA hybridizing region determined by furtherrestriction mapping and Southern analysis. Sequence analysis of the cDNAhad determined that is contains an open reading frame of 552 bases.Presumably the promoter of the gene is located in a Sal I/Eco RIfragment (2.1 Kb). To test this, a chimeric construct was made asfollows. ASK⁺ vector was modified to include a unique Nco I site in thepoly linker. The Sal I/Eco R1 fragment (2.1 Kb) was cloned into SalI/Eco RI site of Nco-SK⁺ vector followed by an Nco I/Xho I GUS fragmentfrom the p2117 plasmid. The resulting plasmid pA11B-2117 was then testedfor GUS activity in hypocotyl tissue. It is found that pA11B-2117expressed GUS. From these results we conclude that A11-B promoter islocated in the Sal I/Eco R1 fragment. Sequence of the A-11 gene promoteris given in SEQ ID NO: 27.

i. Characterization of B8 Gene and Its Promoter

The insert of B8 cDNA clone was used to isolate a phage from Sea Islandgenomic library (19 Kb insert). A Sal I/Bam HI fragment from the phageDNA that hybridized BH8 cDNA insert was subcloned into SK⁺ vector. Thisgenerated a 9.5 Kb fragment that contained the B8 gene. A 2.5 Kb DNApart was then sequenced and was shown to contain homologous sequences toB8. SEQ. ID NO: 28 is the B8 gene. Based on the sequence comparison ofthe cDNA and gene, we determined that a 2.1 Kb Bam HI/BstB1 fragmentwould contain the B8 promoter. In order to create an Nco I site at the3' end of the promoter fragment for convenient cloning of genes, wemodified the B8 promoter by PCR. Two PCR primers MEJ117 and MEJ118, (SEQID NO: 30 and SEQ ID NO: 31) were used to amplify a 120 bp fragment.MEJ118 contained a BstBI and an Nco I site. The PCR product was digestedwith BstBI and purified. Similarly the B8 vector, pSKSIB8 clone was alsodigested with BstBI and a 7 Kb fragment was gel purified and ligated tothe PCR product. Recombinant clones containing the B8 vector were thenscreened and the orientation of the 120 bp insert in B8 was determinedby PCR. Now the plasmid was ready for determination of B8 promoteractivity. The above plasmid was digested with Nco I and Sal I and gelpurified. A GUS-containing clone pSIB8GUS was then introduced intocotton hypocotyl tissue through particle bombardment. Appropriatecontrol experiments were included. These studies show that B8 fragmentcontains promoter activity.

j. Characterization of Sea Island B6 Gene and Its Promoter

A phage, EMBLSIB6-1A, was identified from the screening of the genomiclibrary. A 12.3 Kb Sal I fragment of EMBLSIB6-1A was subcloned into SK⁺vector. After determining that B6 cDNA hybridizes to a 4.3 Kb Xho I/SalI fragment, we isolated the 5' end of the B6 gene as a Bam HI/Xho Ifragment (2.3 Kb) and ligated it into Xho I/Bam HI sites of SK⁺ vector.Subsequently, a 2.0 Kb Xho I/Sal I GUS fragment of p2117 was ligatedinto the B6 vector at the Xho I site. This plasmid pB6-2117 was thenintroduced into hypocotyl tissue along with control plasmids by particlebombardment. GUS activity was detected after 24 hours of bombardmentindicating that the Xho I/Bam HI fragment contains a functionalpromoter. Sequence of this 5' flanking region containing B6 promoter isgiven in SEQ ID NO: 29.

k. Characterization of Sea Island E9 Gene and Its Promoter

cDNA pCKFb23-E9 was used to isolate a phage EMBLSI-E9 from Sea Islandlibrary. The 12.4 Kb Sal I insert from EMBLSI-E9 was subcloned into Sk⁺vector (pSKSI-E9). We determined the DNA fragment that hybridizes to E9cDNA, by restriction mapping and Southern hybridization. The cDNA andthe gene contains an Nco I site at the 5' end. DNA fragment 5' to theunique Nco I site (an Nco I/Xho I fragment of 7.0 Kb) was used togenerate a chimeric GUS plasmid to test promoter function of the NcoI/Xho I fragment. In particle bombardment experiments the pE9-2117construct was found to be active in expressing GUS enzyme.

l. Characterization of Sea Island A12 Gene and Its Promoter

The cDNA was used to isolate two genomic clones, EMBLSI-A12A andEMBLSI-A12B from Sea Island library. The inserts of the phages weresubcloned into SK⁺ vectors. The subclone of EMBLSI-A12A was furthercharacterized in terms of restriction map and Southern analysis. Fromthese studies we concluded that a Sal I fragment (6.7 Kb) contains theA12 gene.

10. Tissue-Specific Expression of Fiber Gene Promoters in TransgenicPlants

In order to demonstrate that various fiber gene promoters can be used toexpress foreign proteins in cotton fiber in a tissue-specific manner,the following experiments were carried out. The coding region of acarrot extensin gene was fused to the E6 promoter (restriction fragmentSal I/Nco I, SEQ ID NO: 21). Extensin is a hydroxyproline richglycoprotein found in the cell-walls of a number of plants. Chem andVarner. EMBO J. 4:2145-2151 (1985). We have ascertained that extensincDNA did not hybridize to control cotton tissue RNAs, including fiber.Thus, extensin is not expressed in cotton plants.

A chimeric extensin gene with a fiber-specific promoter was constructedas follows. A 1.34 Kb Eco RI/Dra I fragment containing the extensin genewas excised from plasmid pDC5A1 (a generous gift from J. Varner's lab),and subcloned into Sma I/Eco RI sites of SK+ vector to generate plasmidSK-Extensin. In order to facilitate further cloning, a Nco I site wasintroduced into the 5' noncoding region of PCR as follows. Two PCRprimers MEJ15 and MEJ16 were used to amplify a 166 bp fragment from thecarrot extensin gene. MEJ16 contained an Nco I and Xho I sites. Theprimer sequences are shown in SEQ ID NO: 32 and 33.

Plasmid SK-Extensin was digested with Xho I/Ava I and the large plasmidfragment purified by gel electrophoresis. The PCR product was digestedwith Xho I/Ava I, gel purified, and subsequently ligated intoSK-Extensin. The coding region and the 5' and 3' noncoding regions ofextensin can be excised as a Nco I/Bst XI (1.51 Kb) fragment from theresulting plasmid. This fragment is ligated into a Nco I/Bst XI digestedE6 promoter vector pSKSIE6-3B (supra). The new plasmid was calledpE6-3B-Ext.

A GUS marker gene was added to the above construct as follows: A 35sviral promoter-driven GUS marker gene was excised from plasmid p2119(supra) as a Xho I/Sal I fragment and was cloned into the Xho I site ofpE6-3B-Ext. The construction of pE6-3B-Ext/GUS is shown in FIG. 10. Theconstruct was ready for transformation into cotton.

More than 30 transgenic cotton plants containing E6-3B-Ext gene weregenerated. A number of these plants were then tested for expression ofextensin in various tissues. By northern analysis we observed expressionof extensin in 10 and 15 day fiber, but not in leaf, ovule, stem, orroot. This result indicates that E6 promoters can be utilized to expressproteins in fiber in a tissue-specific manner. Moreover, the introducedgene is developmentally regulated.

11. Transgenic Cotton Plants Containing H6 Promoter

We have demonstrated that the H6 promoter can be utilized to expressproteins in fiber. An Agrobacterium tumefaciens hormone gene, tryptophanmono-oxygenase (TMO) that takes part in the synthesis of hormone indoleacetic acid (IAA) was fused to H6 promoter and incorporated into cottongenome through particle bombardment (McCabe et al., supra). Examinationof the IAA content from transgenic cotton fiber provides evidence thatthe gene is active in fiber.

12. Assessing Fiber Quality

When the transgenic cotton is able to produce mature fiber, the fibermust be examined in order to determine whether advantageous alterationshave occurred. Cotton fiber length is genetically determined andtherefore varies from cultivar to cultivar. Commercially, Americanupland cottons are classified as either short staple (up to 1 inch; 2.5cm), medium staple (11/32 to 13/32 inch; 2.63-2.78 cm), or long staple(over 11/8 inch; over 2.86 cm). Instruments such as a fibrograph and HVI(high volume instrumentation) systems are used to measure the length ofthe fiber. HVI instruments compute length in terms of "mean" and "upperhalf mean" (UHM) length. The mean is the average length of all thefibers while UHM is the average length of the longer half of the fiberdistribution.

Fiber strength is usually defined as the force required to break abundle of fibers or a single fiber. In HVI testing the breaking force isconverted to "grams force per tex unit." This is the force required tobreak a bundle of fibers that is one tex unit in size. In HVI testingthe strength is given in grams per tex units (grams/tex). Fibers can beclassified as (1) low strength, 19-22 gms/tex, (2) average strength,23-25 gms/tex, (3) high strength, 26-28 gms/tex, and (4) very highstrength, 29-36 gms/tex.

The micronaire reading of fiber is obtained from a porous air flow test.The test is conducted as follows. A weighed sample of cotton iscompressed to a given volume and controlled air flow is passed throughthe sample. The resistance to the air flow is read as micronaire units.The micronaire readings reflects a combination of maturity and fineness.Since the fiber diameter of fibers within a given variety of cotton isfairly consistent, the micronaire index will more likely indicatematurity variation rather than variations in fineness. A micronairereading of 2.6-2.9 is low while 3.0-3.4 is below average, 3.5-4.9 isaverage and 5.0 and up are high. For most textile applications amicronaire of 3.5-4.9 is used. Anything higher than this is notdesirable. Of course, different applications require different fiberproperties. A fiber property that is disadvantageous in one applicationmight be advantageous in another.

It is to be understood that all nucleotide sequence and size datapresented herein is approximate and, although presented as bestunderstood at the present time, may be subject to some variation.

It is to be understood that the present invention is not confined to theparticular construction and arrangement herein illustrated anddescribed, but embraces such modified forms thereof as come within thescope of the following claims.

    __________________________________________________________________________    SEQUENCE LISTING                                                              (1) GENERAL INFORMATION:                                                      (iii) NUMBER OF SEQUENCES: 33                                                 (2) INFORMATION FOR SEQ ID NO:1:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 26 base pairs                                                     (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: Oligonucleotide                                           (iii) HYPOTHETICAL: YES                                                        (iv) ANTI-SENSE: NO                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1:                                       ATGCTGGTACCTTTTTTTTTTTTTTT26                                                  (2) INFORMATION FOR SEQ ID NO:2:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1067 base pairs                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D ) TOPOLOGY: linear                                                         (ii) MOLECULE TYPE: cDNA                                                      (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Gossypium hirsutum                                              (B) STRAIN: Coker 312                                                         (D) DEVELOPMENTAL STAGE: 15 day old fiber cells                               (F) TISSUE TYPE: fiber cells                                                  (vii) IMMEDIATE SOURCE:                                                       (A) LIBRARY: CKFB15A1                                                         (B) CLONE: E6                                                                 ( xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:                                      ACACACACAAGTAAAGCATTAGCAACCATAGCCATGGCTTCCTCACCAAAACTCTTCTCT60                ATGTCTATCCTCTTCCTTTTTGCCCTCTTCTCCATGCAAATCCATGCTAGAGAGTACTTC120               AGCAAATTCCCAAGAGTTAACATCAATGAGAAAGAGA CAACAACCAGAGAGCAAAAGCAC180              GAGACCTTCGTTCCCCAGACCACCCAAAAGCCAGAAGAACAAGAGCCAAGGTTCATTCCT240               GAAACCCAAAATGGTTATGGCCTTTACGGCCACGAGTCAGGCTCAAGCCGGCCCAGTTTC300               ACCACCAAAGAAACCTA TGAACCCTATGTCACCCCTGTTAGATTCCACCCTGATGAGCCC360              TATAACAGCATCCCCGAATCCTCCAACAATAAAGACACTTACTACTACAACAAGAATGCC420               TACGAGTCCACTAAGCAGCAAAACTTGGGCGAGGCCATTTTCACCGAGAAAGGATGGAGC480               ACCAAGGAAAACCAGAACAACAACTACTACAACGGCAACAATGGTTACAACAATGGCGAG540               AAGCAAGGCATGAGCGATACTAGGTACTTGGAGAATGGAAAGTACTACTATGACGTCAAG600               AGTGAGAACAACTATTATCCAAACCGGTTCGACAACTCAAGAGG AGTTGCTTCGAGGAAC660              GAGTTCAATGAGAATCGTTACAACAACATGGGAAGGTACCACCAGAACCAAGAGGAGTTC720               GAGGAAAGCGAGGAAGAGTTCGAACCCTGATCACCTGTCGTACAGTATTTCTACATTTGA780               TGTGTGATTTGTGAAGAACATCAA ACAAAACAAGCACTGGCTTTAATATGATGATAAGTA840              TTATGGTAATTAATTAATTGGCAAAAACAACAATGAAGCTAAAATTTTATTTATTGAGCC900               TTGCGGTTAATTTCTTGTGATGATCTTTTTTTTTATTTTCTAATTATATATAGTTTCCTT960               TGCT TTGAAATGCTAAAGGTTTGAGAGAGTTATGTTCTTTTTCTCTTCCTCTTTCTTTTT1020             TAACTTTATCAAACAATTTTTGAATAAAAATGTGAGTATATTGTAAC1067                           (2) INFORMATION FOR SEQ ID NO:3:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 913 base pairs                                                     (B) TYPE: nucleic acid                                                       (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Gossypium hirsutum                                              (B) STRAIN: Coker 312                                                         (D) DEVELOPMENTAL STAGE: 15 day old fiber cells                               (F) TISSUE TYPE: fiber cells                                                  (vii) IMMEDIATE SOURCE:                                                        (A) LIBRARY: CKFB15Al                                                        (B) CLONE: H6                                                                 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3:                                       TTTCACACGGGTTGTGGCGTAGTTTAAGCAGAGAGGGTGCGCAGGATAAA50                          GCTATTCACCATTGTTTCAACATGAAGGTTTGTAATAAAAATTTGTTTCT100                         ATCAGCATTGCTTTGCATTGCTGT TGCAGGAGTTTTGGGTCAAGCTCCTA150                        GTAATCCTCCTACGTCTACGCCGGCGACACCCACACCACCGGCTTCTACT200                         CCTCCTCCGACGACTCAAGCACCGCCTACACCAACCGCCACTCCGCCACC250                         GGTTTCTACTCCTCCTCCCACTTCATCACCGCCCCCA GTGACAGCTTCTC300                        CACCCCCAGTTTCAACTCCTCCACCCAGTTCTCCTCCTCCTGCAACTCCA350                         CCACCTGCTTCTCCTCCTCCTGCAACTCCACCTCCAGCTTCTCCACCTCC400                         TGCCACTCCTCCACCAGCTTCTCCACCTCCCGCCACTCCACCACCTGCAA 450                        CCCCACCGCCAGCAACTCCTCCTCCTGCTACCCCACCACCAGCTCCATTG500                         GCTTCTCCTCCAGCCACAGTCCCAGCTATCTCTCCAGTACAAACACCATT550                         GACCTCGCCACCAGCTCCGCCGACCGAGGCCCCAGCACCTACCCTCGGGG600                         CTGCTA CGCCAGGTCCAGCTGGAACAGACACGAGCGGAGCAAATCAAATG650                        TGGACCGTACAAAAGATGATGGGAAGCTTAGCCATGGGATGGGCTCTGCT700                         CAATCTGATGGTTTAAAACAACCGTGTGCCTCACATTTGATGCCATAGCT750                         GTGTAATGTTTCATTCAAT TGCTTATTTCGGCCTTGTTTTTCTCGTATTT800                        TATGGGCTGATGTCTCATATGGGACTTTTCTACTATACGTATATGAGAGC850                         CTACATTACTTTACCATTATATTGTATTCTTTGAGACATTATTATTATTT900                         TTTTACCTTTTGA 913                                                             (2) INFORMATION FOR SEQ ID NO:4:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 659 base pairs                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Gossypium hirsutum                                              (B) STRAIN: Coker 312                                                         (D) DEVELOPMENTAL STAGE: 15 day old fiber cells                               (F) TISSUE TYPE: fiber cells                                                  (vii) IMMEDIATE SOURCE:                                                       (A) LIBRARY: CKFB15Al                                                         (B) CLONE: C12                                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4:                                       CAACAATCAGCAATACTCCAAGCAACCATTTTCCTTACAAGTTTGTTTTT50                          CTTG TGATTAATCCATATGGCTAGCTCAATGTCCCTTAAGCTTGCATGTC100                        TGTTGGTGTTGTGCATGGTGGTGGGTGCACCCCTGGCTCAAGGGGCCATA150                         ACCCGTGCTGATGGCTTAGTCGGCCTCCCACGCTGCCTTCCTTTTTTGTC200                         AGGGAATGGTGATGGTG CTGATGCCACAGGTTGCTGTGCCATCGTCATGA250                        ATGCCTTGGGATCGCTCTGTGGTGATACATAGGAACCGATCTAGCTTGAA300                         ATCGGGTTCGGATTTGGGTGGAATTTCAAATTGGTGTGTTATGGAATCCC350                         AACTTAATCGTGTTTAAGGGTGGGATCCAA TTGTGTGATACATTACAGAG400                        CATGGTTGTGGATTGTTTTCTCATATGTTTTGATTGACTTGCTTGCTACA450                         TTGGATGATTTGATAAGGTGACCAGTTTACCTGGGTATCCAACCATCATC500                         GGATTACTTTTTAATAATTTTTTGTTTCTTGTTTATGTTGTC TGCCTTTT550                        TGTTTCTTGATCTATAATATTATATTTGGCCAAATTTCTCATTTTCCAGA600                         TGTAGCTTATATATATATATTCAATAAAGTATATTGGTTTAAAAAAAAAA650                         AAAAAAAAA659                                                                  (2) INFORMATION FOR SEQ ID NO:5:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 690 base pairs                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Gossypium hirsutum                                              (B) STRAIN: Coker 312                                                          (D) DEVELOPMENTAL STAGE: 15 day old fiber cells                              (F) TISSUE TYPE: fiber cells                                                  (vii) IMMEDIATE SOURCE:                                                       (A) LIBRARY: CKFB15Al                                                         (B) CLONE: B8                                                                 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5:                                       CACCAACGGACAATGCTTTCTCCAGCCTTAAATCGGGCACATTGAATTCACTCACCGATG60                AACAAAAAGTGGAGCTGG TGCAATTCCACATCGTCCCAACATACCTCACCTCGTCTCAGT120              TCCAAACCATTAGCAATCCTTTGAGAACCCAAGCTGGTGATAGTGGCGATGGCAAGTTCC180               CTCTCAATATCACCACTTCGGGGAACTCCGTGAATATAACAACAGGGTTGACAAACACCA240                GTGTTTCCGGCACTATTTACACTGATGGTCAGCTTGCTGTTTATCAAATCGATCAAGTTC300              TTCAACCATTGCAAATATTTGCACCTAGGCCTCCAGCTCCAGCACCGGCACCGGCAAAGT360               CGAAGAATAAGAAGGCTACCACCGTTGCTGATAGCCCCGATGTTACCCCA GCTGATAACT420              CCAAAGCGGCCACCTTGCAAAATGTTGGTTTGTTTGGAGTTGCTGCTCTAGTTATTGCAC480               TTTCTTTGTGACCATGAAAATGGAGAAAAGAAGAAGACAGTGATTTTGATGGTGATCAAG540               ATGGCGAGTGTTTTTTATTTTTTCAATAATTA TCATTTAAAAAATTTATGTTCTGTATGA600              ANGATTGAATTTTGAGTTTGTCTTGTTGATTTCATTTATTTTTGTTTTGAAATTTTCTTT660               GTTATCTCTTATTTCTCAATTGTAATTGTG690                                             (2) INFORMATION FOR SEQ ID NO:6:                                               (i) SEQUENCE CHARACTERISTICS:                                                (A) LENGTH: 727 base pairs                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Gossypium hirsutum                                              (B) STRAIN: Coker 312                                                         (D) DEVELOPMENTAL STAGE: 10 day old fiber cells                               (F) TISSUE TYPE: fiber cells                                                  (vii) IMMEDIATE SOURCE:                                                       (A) LIBRARY: CKFB10                                                           (B) CLONE: B12                                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6:                                       ATAACCGTGACAGCCACCAACTTTTGTCCACCTAACTATGCTTTATCTAG50                          TGACAATGGCGGGTGGTGCAATCCCCCACGAGAACACTTTGATT TGGCCG100                        AACCGGCATTCTTGCGGATAGCAGAATATCGAGCTGGAATCGTCCCTGTT150                         ATGTTCAGAAGGGTGTCATGTGTGAAGAAAGGAGGCATCAGGTACACCAT200                         GAATGGACATTCGTACTTCAACATGGTGTTGATAACGAACGTGGGAGGGG250                          CAGGGGATATAACGTCAGTGTCCATCAAGGGTTCCAGAACAGGATGGCTA300                        CCTATGTCCAGAAATTGGGGCCAAAACTGGCAGAGCAATGCTTACCTTAA350                         CGGACAAAGCCTCTCTTTTAAAGTGACTGCCAGCGATGGCAGGACTATCA400                         CAGCCTACAATGT AGTGCCTGCTGGTTGGCAATTCGGACAAACTTTTGAA450                        GGAGGCCAGTTTTAAGACAATATTATAGTGTCTGTCTAATATAAAACTGG500                         AATTGACATATTACTTATATAAGGCACATGAGCGTTTTATGCCGAGGTAG550                         CAAAATGGCGCCCGCTGGCTTTATGT GTGAAATAGGCGAGCAAGTGCCAT600                        TAGCCTATAATCTATACATTTCTTATAGTGAACCAAACTATTAAGTTTGA650                         ACTCTAGAGGATATATCCATAATGTCTGAAATTTGTTTGTTGATGATTGA700                         CCATGATATTTATGCTTTTCATTATTG 727                                               (2) INFORMATION FOR SEQ ID NO:7:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 989 base pairs                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Gossypium hirsutum                                               (B) STRAIN: Coker 312                                                        (D) DEVELOPMENTAL STAGE: 10 day old fiber cells                               (F) TISSUE TYPE: fiber cells                                                  (vii) IMMEDIATE SOURCE:                                                       (A) LIBRARY: CKFB10                                                           (B) CLONE: A11                                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7:                                       TAAAAGGATAGCATCTGCCCTTACAAAGATGAAAATACAAGCAAAATCGA50                          AGACTTCAGAG ATCATCCTTAAAAGTTGAACACAAAGTTAACTTGAAATA100                        CCAGCTAAAGTGATAATAAAACTCGACCATAGAATTTCGGAAACTTCGAA150                         ACATTCACCAAATAAAACCGTCCCTCGAATTTCAACTATCAAACAGTAAG200                         GCTCAACTCACAAAAGCCTTGGAA AGAGGTACACAAATGTTTTATCCTAC250                        TTATTCATTCAATCAATAAAATAAATGGAACATGAACTCCATCCTCCTTG300                         GTTTGACAATACCAGCTTTCACAATTAAGATTCTATACCAGATTCATGAG350                         CTTGAACGGAATCACTCTGAAACAATTACTACATGTA ACAATGGAAACGA400                        AATGGAAAAACAAAAAAAAGTTGGTTTAATTAATTATTAGTTACCCTTGA450                         AGACCTTGGCATTGGTGGAGTAACTCTTGGCATGAAAGTCTGAGAACAAG500                         TAGAGAAGAGAGACGTTGAAGGCTCCATTGAAACACCAACTCAGAATCCC 550                        AGAGCAGCCAGAAGCAGTGAAGTGGTAGAACACAAGCATGGCCATGATCA600                         AAAAGCTTAACCGGAACTGCACCAGTTGAAAATCCGTCACCATTTTCTTC650                         CACTTGGGGTGCATCCCCAGGGTGCACAACAGGTAATAGGAGTACATTAC700                         GACATG CACCACGCAGTTGGTGATCAGCACCATGGGTACGGAGGACTGAG750                        CACTGTCTAAGCAAATATAACACATGATGACCACCATGGAGTGATGGTAG800                         ACGTGAAGGAAGGATAGCCTCTTCATGGATCCGCTGAGGATGATCAAAAG850                         GGTGTCCATGAATTCAACA ATCTTGGAGAGGTAGAAGATGTATGCCCAGA900                        AAAAGAGAGGGCCCGATGGGGATGTACCCCTAGGGAAGCAAACGAGGGTG950                         TTGAAGTTAGGCACCTGGGAGAAGATGGAAACGAGGCAA989                                    (2) INFORMATION FOR SEQ ID NO:8:                                              (i) SEQUENCE CHARACTERISTICS:                                                  (A) LENGTH: 498 base pairs                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Gossypium hirsutum                                              (B) STRAIN: Coker 312                                                         (D) DEVELOPMENTAL STAGE: 10 day old fiber cells                               (F) TISSUE TYPE: fiber cells                                                  (vii) IMMEDIATE SOURCE:                                                       (A) LIBRARY: CKFB10                                                           (B) CLONE: D7                                                                 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8:                                       CAATATTTGCTGTGGCCATGTTTGTTGTGTCCGGGACCATGGCACAGGATATTGCTCCTT60                CTCCTGCAATGGCTACCGGAGCAGGCTCTGCTTTGCCGGTTTCCGCTGT CTTCTTATGCT120              CTTCCATGTTGGTCTCTTTAATTGCTCTCTTGGTGCATTGAATTCAAAGCTTTTCAAGAC180               TTTATGACATTGGCTACCCTTAATTTCACTCTCACTGGTGATGAGGGGAGTAGCCTCTAA240               TCTTCTCCGAGATAATATTTGGGTGTTAT CAATTTTCAATTTCTCTAAAGTTTTAAATAT300              CTCTATATATATCATGTTACATTAGTGACATAAATTTGATTTTGTAATGTAATTGGTGTG360               ATTTTCTTATATATCATAGATATATAAGGGTATTGATCCTATTTCTTTTGTATTCATATT420               ATGCAGTTT GCTTTGCTTTGCCTTTTGTAATAATAATTTTTTTTTGGGTGGTTGAAAAAA480              AAAAAAAAAAAAAAAAAA498                                                         (2) INFORMATION FOR SEQ ID NO:9:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 668 base pairs                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Gossypium hirsutum                                              (B) STRAIN: Coker 312                                                         (D) DEVELOPMENTAL STAGE: 10 day old fiber cells                               (F) TISSUE TYPE: fiber cells                                                  (vii) IMMEDIATE SOURCE:                                                        (A) LIBRARY: CKFB10                                                          (B) CLONE: C2                                                                 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9:                                       TTTTTTAAACAAAAGCATAGTAGTTTTTATACCCCACTGGTGAAGAATGA50                          CAAACACAGTTTCCAAATTCAAATGCATCATCTAAACAAATATGCCCCTT100                         TCTTAGTCTTATCTGCATGGTTTTGCTTG CTGGAAATGAAAAAGCAAAAA150                        TGAAAGAAAAAGAAAAAAGGTGAAAACAACCTTCAAGGTTTAAGAGATGA200                         TATGTAATTTTTCACTTTTTTCAACGCATTGCAGCAAGAGGGTTCCTTTT250                         CCATTGCAGAGGCTGATATGTCTTCTCTGTTTCCTCTATTT TAGTCCATG300                        GTAATTTGTGTTTAGCCACCTTTCGCTTCCTAGCTGATACTCCCAGATAG350                         TCTCCAGCGTTCTTGAGGCAGAGTCCTTCTTGGACATCACAAATGGGGTA400                         ATCACTAGGGCAGCAGTATTCAGTTCCAGTACAGCAAACAGCATTTTCAT450                         ATTCACAGCAGCCGTATATTAGGCAATAATCATAGAATTCAAAAAGGCAA500                         CAGCATGTCTCATCACTTGAACAATAGGAAAAGTCTCCACAATCACTTGG550                         TGAAGGAGATGGAGGTGGTGGTGGTGGTGGAGTTGAAGGAGGTGGCGGTG600                         GAGGAACACT TGGCGATGGATAAGGGGATGGTGAAGAGGATTGTTTAGTT650                        GGATAAGAAGCCATGGCA668                                                         (2) INFORMATION FOR SEQ ID NO:10:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 570 base pairs                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                       (D) TOPOLOGY: linear                                                         (ii) MOLECULE TYPE: cDNA                                                      (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Gossypium hirsutum                                              (B) STRAIN: Coker 312                                                         (D) DEVELOPMENTAL STAGE: 10 day old fiber cells                               (F) TISSUE TYPE: fiber cells                                                  (vii) IMMEDIATE SOURCE:                                                       (A) LIBRARY: CKFB10                                                           (B) CLONE: C12                                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10:                                      TATCAACTACCAACCACCCACTGTTGTCCCTGGTGGTGATCTCGCCAAGGTCCAGAGAGC60                TGTTTGCATGATTTCTAACTCCACCAGTGTTGCTGAAGTCTTCTCTCGCATTGACCATAA120               GTTTGATCTTATGTACGCCAAACGTGCCTTC GTTCACTGGTATGTTGGTGAGGGTATGGA180              AGAAGGAGAATTCTCAGAGGCTCGTGAGGATCTTGCTGCATTGGAGAAGGATTATGAAGA240               AGTTGGTGCTGAGTCAGCTGAGGGTGATGAGGGTGAAGATGATGAGTACTAAGAAGTACG300               TGTGATTGAGTGTG CTTACGATGTCTTTTATTACCTGTTGCTTCCGTGTTATGCAAGCTT360              CTATTCTTTGAAGCTTGTTAAAGACTTGAAATGGTTTACTGTATGTTATGCTGTTTTATG420               TTTTATTTGTGTTTGGGTTGAAACTTGATATTTTTGTTGGGTGTATTTGAAAGTAAATTG480               TTAAGGGAACTTGTGAATGCTCAACAGGTATAAAAAAAAAAAAAAAAAAAAAAAAAAAAA540               AAAAAAAAAAAAAAAAAAAAAAAAAAAAAA570                                             (2) INFORMATION FOR SEQ ID NO:11:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 432 base pairs                                                     (B) TYPE: nucleic acid                                                       (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Gossypium hirsutum                                              (B) STRAIN: Coker 312                                                         (D) DEVELOPMENTAL STAGE: 10 day old fiber cells                               (F) TISSUE TYPE: fiber cells                                                  (vii) IMMEDIATE SOURCE:                                                       (A) LIBRARY: CKFB10                                                           (B) CLONE: C1                                                                 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11:                                      TTTCATTTCACTATATACACACACACATACACAAACCAAGCAACCATGGA50                          TACAAGAAGCAAAACACCTAATGAACCATAGCTGCTTTATTCTAATAGTA100                         AAAACCCAAATACATCATA TATTATTTAGATCAGCTTCCATAATATGCTT150                        AGCTTTTTTTTCTCATTTACAATTGCAAGGGTTGCAAGTGCAGTTATCTC200                         CACATTTGCAGCCATTTTCAGCCCCAGTTTCCATTTCAGCTCCATCAAAG250                         TGCACTTTCCGGGGTGCCACGCCAAGAACAA GTGTCCCGGTTGTGGTTTG300                        CTCAGCAAAGTTCATCTCAGGGTACATCTTGCAACCGCCGCAGCCGCTGC350                         CGCACTTGCAACCGGAGCCGCAACCGCAGTTTCCACCACAGCAAGACATT400                         TTTCTTCACCTCACTGATCACTAAAGGGCGAT 432                                          (2) INFORMATION FOR SEQ ID NO:12:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 320 base pairs                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Gossypium hirsutum                                               (B) STRAIN: Coker 312                                                        (D) DEVELOPMENTAL STAGE: 10 day old fiber cells                               (F) TISSUE TYPE: fiber cells                                                  (vii) IMMEDIATE SOURCE:                                                       (A) LIBRARY: CKFB10                                                           (B) CLONE: A8                                                                 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12:                                      CAATCAAATTAACTTAAAAGAAGTGATATTCAGGATACAAGGCCAATGCCAAACAGATTC60                AACACAA ACATAAGCTGCATACATATAGGCAACTTGAAGTTATTGAATAGACGGGATTCA120              GAGAACAAAGCTGGTTAAAACATGAGGTACGATACAGATATTACATTGGCATGTTCTTCA180               GAAGATTTTTTTGGATGGCTAAGTGGAACCACCAATTTTGGGATGATCTGGAGGTG AGGG240              TTTTGGGATGTAGGGAATAAGAGGAGTACCATGCTCAATAGGACCAGGTTTTATGGATGC300               TTTGGAGCTTGGAACTGGAT320                                                       (2) INFORMATION FOR SEQ ID NO:13:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 399 base pairs                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Gossypium hirsutum                                              (B) STRAIN: Coker 312                                                         (D) DEVELOPMENTAL STAGE: 10 day old fiber cells                               (F) TISSUE TYPE: fiber cells                                                   (vii) IMMEDIATE SOURCE:                                                      (A) LIBRARY: CKFB10                                                           (B) CLONE: A9                                                                 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13:                                      TCAACTCCGCCGCCCAAACAACACCAGACCGGCAAGCAGCTTGCAAATGCATCAAAAGTG60                CGGCCGCCGGCATTTCTGGCATCAACTATGGTATTGCAAGCGGACTCCCAGGCAAG TGCG120              GTGTCAACATCCCTTACAAGATCAGCCCTAGCACTGACTGCAACAGCGTCAAGTGAAGTT180               TTGGCATGGAAAGTTCACCAGCTAGTGGAAGCCAAAATAACGATAGCTACAGAATAAATA240               TGGATGTTAAAATTCCAGAGTTGTGGGTTGTGTACT ATGCCGCTTTATGCGACTACGTAA300              TATTAACTTTATCTACAAATTAATATCACTCGTCTCCATTTCCCATTTTAAAAAAAAAAA360               AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA399                                    (2) INFORMATION FOR SEQ ID NO:14:                                              (i) SEQUENCE CHARACTERISTICS:                                                (A) LENGTH: 455 base pairs                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Gossypium hirsutum                                              (B) STRAIN: Coker 312                                                         (D) DEVELOPMENTAL STAGE: 10 day old fiber cells                               (F) TISSUE TYPE: fiber cells                                                  (vii) IMMEDIATE SOURCE:                                                       (A) LIBRARY: CKFB10                                                           (B) CLONE: D4                                                                 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14:                                      AGCAGCTGTGCCTAGTCCTTCTACTATGCCTAGAGCTTGGACTTTCTTCC50                          TACTCGATCAGATTCTAACATACGTAATCTTGGGAGCTGCTGCT GTTTCA100                        ACCGAGGTGCTTTACTTAGCAAACAAAGGAGACTCAGCCATCACTTGGAG150                         TGCAGCTTGTGGGACATTTGCTGGTTTCTGTCATAAAGCCACAATAGCCG200                         TGGTGATCACGTTTGTTGCAGTCATTTGTTATGCGGTGCTATCACTGGTC250                          TCTTCTTATAGACTTTTCACCAAGTTTGATGCCCCAGTGAACTACCCCAG300                        TAAGACCATAGAAGCTACTGTTTTCCATGGTTGATTTATGTTATTACTGA350                         AATTAATTTACCTTATATTTTCATGTTCTGCTTGTAATAATAATAAAAAA400                         GGTTGCTTACAGT GTGTTTATGTTATATGATTAAATAGAGGTGTTGTCTT450                        TGGTG455                                                                      (2) INFORMATION FOR SEQ ID NO:15:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1080 base pairs                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                       (D) TOPOLOGY: linear                                                         (ii) MOLECULE TYPE: cDNA                                                      (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Gossypium hirsutum                                              (B) STRAIN: Coker 312                                                         (D) DEVELOPMENTAL STAGE: 10 day old fiber cells                               (F) TISSUE TYPE: fiber cells                                                  (vii) IMMEDIATE SOURCE:                                                       (A) LIBRARY: CKFB10                                                           (B) CLONE: B6                                                                  (xi) SEQUENCE DESCRIPTION: SEQ ID NO:15:                                     ACTACTCAACTTTCTCTCTGAATTCCTCCAAGTTAGGAGTTTGGAGAGTG50                          GTCACCGCTGAAGCAAAACAAATTTCTTGGGGGAAAAGAAAATGGAGTTT100                         TCCATGATTTTTATGATTAGCTTCTCTGTATTGATTTTGTGCTCCTCACT1 50                        GGCATATGGTCAAGTTGCAATGAGCACAAACCCGACACCGTCACCCTCAC200                         CAGCACCGGCACCGACACCGGCATACACAAATATCAAAGACTTACTCTCT250                         GTGGCAGGTCCATATCACAAGTTCCTGGGCTACCTCGAGTCGACTAAATT300                         AATCG ACACGTTCCAAATCCAAGCCAACAACACGGTTGAAGGCATTACGA350                        TTTTCGTACCGAAAGACAGCGCATTCAAGGCTCTTACGAAGCCTTCATTG400                         TCAAATCTAACTGATGATCAGTTCAAATCAGTGCTCCTTTACCATGCCTT450                         GCCACGATACTATGC CCTTGCGGACTTCAATGACCTAAGTGAGAAAGGCC500                        CTATTAGTACACTTGCTGGTGGCCAATACACTTTGCAATTCAACGATGAG550                         TCTGGTACCGTCCGCCTCGATTCCGGATGGAGCAAAACAAAAGTCACTAG600                         CGCAGTACATACGTCCAAGCCAGTC GCAGTCTATCAAATCGATAAGGTCC650                        TTCTTCCGGAGGCCATTTTCGGGACCGACATACCTCCGACACCTGCACCT700                         GCCCCGGCTCTTGGTATTGGCCCATCAGCTGACACTCCATCAGCAGCAAA750                         ATCCGAAGAAACTGGTTCCTCATCAAAGCCTTCGT TTTCGGGTTCATCAT800                        CTCCTAGGATCATGATGAACTCGGGCATTTGGACTCAGCTGGTTTTGGCA850                         TTCTTAGGTGGATGGCTGGTTCTCTTTTTCTGAGACGTTATAATTTTATG900                         TTGAAAGGGGGGCACATATGGGGTTCTCAATTTTCTGTGATTTTT AGACC950                        CCATTTTCTTTCATATATATGTTACTGTGTGTTATTATAAAAAGAATGTT1000                        ATTGTGTGTTAAGAATATGGTTGTGTTATAATTACCATTTCAATTTTAAT1050                        GGAGTTTTGCCTTAAAAAAAAAAAAAAAAA1080                                            (2) INFORMATION FOR SEQ ID NO:16:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 868 base pairs                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Gossypium hirsutum                                              (B) STRAIN: Coker 312                                                          (D) DEVELOPMENTAL STAGE: 10 day old fiber cells                              (F) TISSUE TYPE: fiber cells                                                  (vii) IMMEDIATE SOURCE:                                                       (A) LIBRARY: CKFB10                                                           (B) CLONE: A12                                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16:                                      CAACGATACTTCAAATACTAATACATTTATCACCAAACCATTGTGATACA50                          GAAAGTAATATCATTTTATCCATTAC CAAGAACATGATTCATGGATAGAT100                        TTCACATAACTAAACTCGTGGGTATATTATTATAGAACAAACTGGATATT150                         GCTTTAAGCTGTTTTAATGCACACTATGATGAAACTTATAGTGTATGAAT200                         CTACTCTTCAGGATTTTTACTTAGGGACCCAAACAATGT GATCCCGAGGA250                        AGGAAATGGCAGACAGGAATGGTTCCTGGTTCAACTTTTAGGACTTGAAA300                         AGCCAAATGCTTAGGGTTCCATGCTGATGTATCTGTGTGGCAGACTGCTA350                         CTGCTTTGGCTTTTGTTCCGTCAGCACCCTCTAAAGGAACCATGTAAGCC4 00                        CTTGTTGTTTCTGATTTATGGCAATAGAAGACAGCATATGCATAATTCTG450                         CTTGTGGCACACTACGGCTTTGTCATCTGTCATCTTCTGCACTCCAGCTG500                         CTATTGTATACTTTTGCATTGGGGTTTGTTTTTCCACTTCTGTTGAGACT550                         GCCTGATC AACTTTCCCTAGTTTGGAAATGCTATAGTCAATCATTGACTC600                        CAGTGAGGTTGCACAATATTTTTCCTCTCCTTCAATCGCTGGCTGTTCGC650                         ACTCCTTAATTGTGTTCTTCATCATCTCTGCCTTCAGTGATCCAGGTTTC700                         ACTGAAAACTTGTTGAAAAT TTCTGGCAACTTGTCAGATGAAAACGGTAT750                        TTTTTGGGCAGTTTGATAAGGTAAGAAAGCTGATTTCTCTGTATTTTCAG800                         TGAAATGCAGGCTCATTGTTGCCCCGGGGTGCATATCCTTTTCCAGAAAG850                         AAAAGAGCCACATTCGGG 868                                                        (2) INFORMATION FOR SEQ ID NO:17:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1283 base pairs                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Gossypium hirsutum                                              (B) STRAIN: Coker 312                                                         (D) DEVELOPMENTAL STAGE: 15 day old fiber cells                               (F) TISSUE TYPE: fiber cells                                                  (vii) IMMEDIATE SOURCE:                                                       (A) LIBRARY: CKFB15                                                           (B) CLONE: E9                                                                 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17:                                      ACACACAAATACACTAAAAATTCTTTGCTTTCTATTTTGTAAACCATGGC50                          TCAT AACTTTTGTCATCCTTTCTTCCTTTTCCAACTTTTACTCATTACTG100                        TCTCACTAATAATCGGTAGTCACACCGTCTCGTCAGCGGCTCGACATTTA150                         TTCCAGACACAAACAACCTCATCAGAGCTGCCACAATTGGCTTCAAAATA200                         CGAAAAGCACAAAG AGTCTGAATACAAACAACCAAAATATCACGAAAAGT250                        ACCCAAAACATGAGAAGCCTAAAATGCACAAGGAGGAAAAACAAAAACCC300                         TGCAAACATCATGAAGAGTACCACGAGTCACGCGAATCGAAGGAGCACGA350                         AGAGTACGATAAAGAAAAACCCGA TTTCCCCAAATGGGAAAAGCCTAAAG400                        AGCACAAGAAACACGAAGTTGAATATCCGAAAATACCCGAGTACAAGGAC450                         AAACAAGATGAGGATAAGGAACATAAAAATGAAGAGTACCATGAATCACG500                         CGAATCGAAGGAGCACGAAGAATACGAGAAAGAA AAACCCGAGTTCCCCA550                        AATGGGAAAAGCCTAAAGAGCACGAGAAACACGAAGTCGAATATCCGAAA600                         ATACCCGAGTACAAGGAAAAGCAAGATAAGAGTAAGGAACATAAAGATGA650                         AGAGTGCCACGAGTCACACGAATCGAAAGATCACGAAGAGTACG AGAAAG700                        AAAAACCCAATTTCTTCAAATGGGAAAAGCCTAAAGAGCACGAGAAACAT750                         AAAGCCGAATATCCAAAAATACCCGAGTGCAAGGAAAAACAAGATGAGGA800                         TAAGGAAGATAAACATGAGTTCCCAAAGCATGAAAAAGAAGAGGAGAAGA850                         AACCTGAGAAAGGCAGAGTACCCTGAGTGGGTTAAAATGCCTGAATGGCC900                         GAAGTCCATGTTTACTCAGTCTGGCTCGAGCATTAAGCCTTAAGCCATAT950                         GACACTGGTGCATGTGCCATCATCATGCAGTAATTTCATGGGATATCGTA1000                        ATTATAT TGTTAATAAAAAAGATGGTGAGTGGGAAATGTGTGTGTGCATT1050                       CATCCATGTAGCAATGCTGAATCTCTTTGCATGCATAGAGATTCTGAATG1100                        GTTATAGTTTATGTTATATCGTTTGTTCTAGTGAAATTAATTTTGAATGT1150                        TGTATCTAATGTTAACA TCACTTGGCTTGATTTATGTTTTAATGAAGTTT1200                       ATGTTGTGTATTTTACTTTAATGATATTCCATGTATTGTTAATTTAAAAA1250                        AAAAAAAAAAAAAAAAAAAAGGCCGAATTGGCA1283                                         (2) INFORMATION FOR SEQ ID NO:18:                                             (i) SEQUENCE CHARACTERISTICS:                                                  (A) LENGTH: 878 base pairs                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Gossypium barbadense                                            (B) STRAIN: Sea Island                                                        (vii) IMMEDIATE SOURCE:                                                       (A) LIBRARY: SIFB10                                                            (B) CLONE: H8                                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:18:                                      AAAAAGGGGAAGAGCTAATGGAAAAAAGTGAAAAGGGTAATGGTTTTGCTCCTGCTACAA60                GGTCTCCAATGGCTTTAATGGGGTCATCTAGAAATGAAAACCAAGAAGTGAATACCAGCA120               TGAGAACTGCCGAGAC CATGCTGCGGTTGGTGCCTATGGCTTTAGGGGTTGCTGCACTTG180              TTGTCATGCTCAAAAACTCACAGTCCAATGACTTTGGCTCCGTTTCATACTCAGATCTTG240               GTGCTTTCAGGTACTTGGTGCATGCTAATGGTATTTGTGCAGGCTATTCCCTTCTTTCAG30 0              CTATTATAGCAGCTGTGCCTCGTCCTTCTACTATGCCTAGAGCTTGGACTTTCTTCCTAC360               TCGATCAGATTCTAACATACGTAATCTTGGGAGCTGCTGCTGTTTCAACCGAGGTGCTTT420               ACTTAGCAAACAAAGGAGACTCAGCCATCACTTGGAGTGCAGC TTGTGGGACATTTGCTG480              GTTTCTGTCATAAAGCCACAATAGCCGTGGTGATCACGTTTGTTGCAGTCATTTGTTATG540               CGGTGCTATCACTGGTCTCTTCTTATAGACTTTTCACCAAGTTTGATGCCCCAGTGAACT600               ACCACAGTAAGACCATAGAAGCT ACTGTTTTCCATGGTTGATTTATGTTATTACTGAAAT660              TAATTTACCTTATATTTTCACGTTCTGCTTGTAATAATAATAAAAAAGGTTGCTTACAGT720               GTGTTTATGTTATATGATTAAATAGAGGTGTTGTCTTTGGTGCTTCTTTTGTAATCTTCA780               GAC TGCTTACTAGAATCCCTTTTAGGTTGCATCATTGGTATCATGTAATGTCAAAAATGA840              AAAAAGATTTATAAGTGACAGCACAAAATGCAAAAAAA878                                     (2) INFORMATION FOR SEQ ID NO:19:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1603 base pairs                                                    (B) TYPE: nucleic acid                                                       (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Gossypium barbadense                                            (B) STRAIN: Sea Island                                                        (vii) IMMEDIATE SOURCE:                                                       (A) LIBRARY: SIFB10                                                           (B) CLONE: H4                                                                 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:19:                                      ATCTCATATTCCGGAAGGCGTCTTGATACACGCCCTCATTTATTTTATTTTTCAAACAAA60                TTTTCCCTTCAGTTTTCGTATAATTTCCTAGGAAAAGAAAAATGAGAGAGTGCATTTCAG120               TTCACATTGGTCAGGCCGGTATTCAGGTCGGAAATGCTTG CTGGGAACTTAGTGTCTCGA180              GCATGGCATTCAGCCTGATGGCCAAATGCCAAGTGATAAGACTGTTGGTGGAGGAGACGA240               TGCTTTCAACACCTTTTTCAGCGAAACTGGTGCCGGGAAGCACGTCCCTCGCGCCATCTT300               TGTTGATCTGGAGCCTACTG TTATCGATGAAGTGAGGACTGGTACGTACCGCCAGTTGTT360              CCACCCTGAGCAACTCATCAGTGGCAAAGAAGATGCTGCCAACAATTTCGCTCGTGGCCA420               TTATACAATTGGCAAAGAGATTGTTGATCTCTGCTTGGATCGTATCCGAAAACTTGCTGA480                TAACTGTACTGGGCTCCAAGGCTTCTTGGTTTTCAATGCTGTTGGTGGTGGTACTGGTTC540              TGGTCTTGGATCTCTTCTCTTGGAGCGTCTCTCTGTTGACTATGGAAAGAAGTCGAAGCT600               TGGTTTCACTGTCTATCCTTCACCTCAGGTTTCTACATCAGTTGTAG AGCCTTACAACAG660              TGTGCTGTCCACTCATTCGCTCCTTGAGCACACTGATGTCGCTGTTCTTCTGGATAACGA720               AGCAATATATGACATCTGCAGCTGCAGGCGTTCTTTGGACATTGAACGACCCACTTATAC780               CAATCTTAACCGCCTTGTCTCTCAGGT TATCTCATCTCTTACCGCATCTTTGAGGTTTGA840              TGGAGCCCTGAATGTGGATGTGACTGAGTTCCAGACTAACCTGGTCCCATATCCCAGGAT900               CCACTTTATGCTTTCTTCTTATGCCCCTGTCATTTCAGCTGAGAAGGCTTACCATGAGCA960               GCTATCG GTGGCTGAGATAACCAACAGTGCTTTTGAACCCTCTTCAATGATGGCCAAATG1020             TGACCCACGACACGGGAAGTACATGGCTTGCTGCCTTATGTACCGAGGAGATGTTGTGCC1080              CAAAGATGTGAATGCGGCTGTGGCCACCATTAAGACCAAACGCACAATCCAATT TGTGGA1140             TTGGTGCCCTACTGGATTCAAGTGCGGTATCAACTACCAGCCACCAACTGTTGTTCCAGG1200              AGGGGACCTTGCCAAGGTTCAGAGAGCTGTCTGCATGATCTCCAACTCAACCAGCGTTGC1260              GGAAGTGTTTTCCCGCATTGATCATAAATTTGAT CTCATGTATGCCAAGCGTGCATTTGT1320             GCACTGGTATGTTGGTGAGGGCATGGAGGAAGGAGAGTTTTCTGAGGCACGTGAAGATCC1380              TCGCTGCACTGGAAAAAGATTATGAAGAAGTTGGCGCTGAGTCTGGTGAAGGAGACGAAG1440              GGGATGAGGAGGAG TATTGAGGGATGTACCGTTATCCGTTTGCTACTGTGATGTATTTTC1500             TTGTTGATTTTGGATATGTGTTTTCAGGGTTGAATACCTCGGATGATGTACTAGTTTTTG1560              GTATTTTCTATGAATAAAGTGCACGGTAAAATTATAAAAAAAA1 603                              (2) INFORMATION FOR SEQ ID NO:20:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1647 base pairs                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: DNA (genomic)                                             (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Gossypium barbadense                                            (B) STRAIN: Sea Island                                                        (vii) IMMEDIATE SOURCE:                                                       (A) LIBRARY: EMBL-SI                                                          (B) CLONE: E62AH3                                                             (xi) SEQUENCE DESCRIPTION: SEQ ID NO:20:                                      ATTTAACAAATATATTTTGAAAAATTGATAAAAATACTAAATGAGGTTTTGGTTGAATAG60                TAAGATATAATTATTACAAATTATAAATATGTAGGTTCAAAATCTATCAT GTGTATATTT120              GTACTATTATTCTATATAAATTGATAACCTTATAAAAGTATCTAATTTAGTTTATGGTTG180               ATTGATCGATAATACCAAATTTATTAAAAATTAATATTAGTAAAGATATATAGTACAAAA240               CTAAACATAAAATTTTATATGTTAAGGAAA TAGCGGAAAAAATATCATATTTGTAGAACT300              GTTTAGCAGTGTGGGAGAATGGGATCATTACAAGGAAAAATGAAATATATATCATTAATA360               CCAAACATAAAAGAAAGCGTCTTTTGATAAAGTTGTTATTGGTGTAATGTGAAGGGACCA420               CAATCATCAC CATTCACCACTTGCTCCTAATTGAGTTGAAATCTTTTTACAACATAGAAA480              ACTAGAAGATCGCCCTTTCTTGCTTCATATATATAGATTTTGTATCATCGCAATTTCACA540               TCACACACACAAGTAAAGCATTAGCAACCATAGCCATGGCTTCCTCACCAAAACTCT TCT600              CTATGTCTATCCTCTTCCTTTTTGCCCTCTTCTCCATGCAAATCCATGCTAGAGAGTACT660               TCAGCAAATTCCCAAGAGTTAACATCAATGAGAAAGAGACAACAACCAGAGAGCAAAAGC720               ACGAGACCTTCGTTCCCCAGACCACCCAAAAGCCAGA AGAACAAGAGCCAAGGTTCATTC780              CTGAAACCCAAAATGGTTATGGCCTTTACGGCCACGAGTCAGGCTCAGGCTCAGGCTCAG840               GCTCAGGCTCAAGCCGGCCCAGTTTCACCACCAAAGAAACCTATGAACCCTATGTCACCC900               CTGTTAGATTCCACCCT GATGAGCCCTATAACAGCATCCCCGAATCCTCCAACAATAAAG960              ACACTTACTACTACAACAAGAATGCCTACGAGTCCACTAAGCAGCAAAACTTGGGCGAGG1020              CCATTTTCACCGAGAAAGGATGGAGCACCAAGGAAAACCAGAACAACAACTACTACAACG1080              GCAACAATGGTTACAACAATGGCGAGAAGCAAGGCATGAGCGATACTAGGTACTTGGAGA1140              ATGGAAAGTACTACTATGACGTCAAGAGTGAGAACAACTATTATCCAAACCGGTTCGACA1200              ACTCAAGAGGAGTTGCTTCGAGGAACGAGTTCAATGAGAATCGT TACAACAACATGGGAA1260             GGTACCACCAGAACCAAGAGGAGTTCGAGGAAAGCGAGGAAGAGTTCGAACCCTGATCTC1320              ACCTTCAGATGATTTCGCTGAAGCAGATCCAAGATGGTTTGGCATCCTTGTGTTTACAAG1380              GAGCAAATCGAAGATATAGCTGAT TTGGCTTTCACGTGATTACGATGAAGCAAATCTAAG1440             ATGATTTGTCGTCTCTGTATCGTTAGAGAACGAATCGAAGTTTGGCATCTTCACTTTGAT1500              GGAGAGCAGACACATAGCAGATCTCACCTTCAGATGATTTCGCTGAAGCAGATCCAAGAT1560              GGTT TGGCATCCTTGTGTTTACAAGGAAAAAATCGAAGACATAGCTGATTTGGTTCATGT1620             GATTACGATGAAGCAAATCTAAGATGA1647                                               (2) INFORMATION FOR SEQ ID NO:21:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1668 base pairs                                                    (B) TYPE: nucleic acid                                                       (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: DNA (genomic)                                             (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Gossypium barbadense                                            (B) STRAIN: Sea Island                                                        (vii) IMMEDIATE SOURCE:                                                       (A) LIBRARY: EMBL-SI                                                          (B) CLONE: E6-3B                                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:21:                                      AAATTATAGCATACCTCACGATGTGGGTGAAGTAAAATTATTTAACAAATATATTTTGAA60                AAATTGATAAAAATACTAAATGAGGTTTTGGTTGAATAGTAAGATATAATTATTACAAAT120               TATAAATATGTAGGTTCAAAATCTATCATGTGTATATTTG TACTATTATTCTATATAAAT180              TGATAACCTTATAAAAGTATCTAATTTAGTTTATGGTTGATTGATCGATAATACCAAATT240               TATTAAAAATTAATATTAGTAAAGATATATAGTACAAAACTAAACATAAAATTTTATATG300               TTAAGGAAATAGCGGAAAAA ATATCATATTTGTAGAACTGTTTAGCAGTGTGGGAGAATG360              GGATCATTACAAGGAAAAATGAAATATATATCATTAATACCAAACATAAAAGAAAGCGTC420               TTTTGATAAAGTTGTTATTGGTGTAATGTGAAGGGACCACAATCATCACCATTCACCACT480                TGCTCCTAATTGAGTTGAAATCTTTTTACAACATAGAAAACTAGAAGATCGCCCTTTCTT540              GCTTCATATATATAGATTTTGTATCATCGCAATTTCACATCACACACACAAGTAAAGCAT600               TAGCAACCATAGCCATGGCTTCCTCACCAAAACTCTTCTCTATGTCT ATCCTCTTCCTTT660              TTGCCCTCTTCTCCATGCAAATCCATGCTAGAGAGTACTTCAGCAAATTCCCAAGAGTTA720               ACATCAATGAGAAAGAGACAACAACCAGAGAGCAAAAGCACGAGACCTTCGTTCCCCAGA780               CCACCCAAAAGCCAGAAGAACAAGAGC CAAGGTTCATTCCTGAAACCCAAAATGGTTATG840              GCCTTTACGGCCACGAGTCAGGCTCAGGCTCAGGCTCAGGCTCAGGCTCAAGCCGGCCCA900               GTTTCACCACCAAAGAAACCTATGAACCCTATGTCACCCCTGTTAGATTCCACCCTGATG960               AGCCCTA TAACAGCATCCCCGAATCCTCCAACAATAAAGACACTTACTACTACAACAAGA1020             ATGCCTACGAGTCCACTAAGCAGCAAAACTTGGGCGAGGCCATTTTCACCGAGAAAGGAT1080              GGAGCACCAAGGAAAACCAGAACAACAACTACTACAACGGCAACAATGGTTACA ACAATG1140             GCGAGAAGCAAGGCATGAGCGATACTAGGTACTTGGAGAATGGAAAGTACTACTATGACG1200              TCAAGAGTGAGAACAACTATTATCCAAACCGGTTCGACAACTCAAGAGGAGTTGCTTCGA1260              GGAACGAGTTCAATGAGAATCGTTACAACAACAT GGGAAGGTACCACCAGAACCAAGAGG1320             AGTTCGAGGAAAGCGAGGAAGAGTTCGAACCCTGATCACCTGTCGTACAGTATTTCTACA1380              TTTGATGTGTGATTTGTGAAGAACATCAAACAAAACAAGCACTGGCTTTAATATGATGAT1440              AAGTATTATGGTAA TTAATTAATTGGCAAAAACAACAATGAAGCTAAAATTTTATTTATT1500             GAGCCTTGCGGTTAATTTCTTGTGATGATCTTTTTTTTTATTTTCTAATTATATATAGTT1560              TCCTTTGCTTTGAAATGCTAAAGGTTTGAGAGAGTTATTGCTCTTTTTTTCTTCCTCTTT1 620             CTTTTTTAACTTTATCATACAAATTTTGAATAAAAATGTGAGTACATT1668                          (2) INFORMATION FOR SEQ ID NO:22:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1618 base pairs                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: DNA (genomic)                                              (iii) HYPOTHETICAL: NO                                                       (iv) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Gossypium hirsutum                                              (B) STRAIN: Coker 312                                                         (vii) IMMEDIATE SOURCE:                                                       (A) LIBRARY: Lambda DASH-CK                                                   (B) CLONE: CKE6-1A                                                            (xi) SEQUENCE DESCRIPTION: SEQ ID NO:22:                                      CTCACGATGTGGGTGAAGTAAAATTATTTAACAAATATATTTTGAAAAA TTGATAAAAAT60               ACTAAATGAGGTTTTGGTTGAATAGTAAGATATAATTATTACAAATTATAAATATGTAGG120               TTCAAAATCTATCATGTGTATATTTGTACTATTATTCTATATAAATTGATAACCTTATAA180               AAGTATCTAATTTAGTTTATGGTTGATTG ATCGATAATACCAAATTTATTAAAAATTAAT240              ATTAGTAAAGATATATAGTACAAAACTAAACATAAAATTTTATATGTTAAGGAAATAGCG300               GAAAAAATATCATATTTGTAGAACTGTTTAGCAGTGTGGGAGAATGGGATCATTACAAGG360               AAAAATGAA ATATATATCATTAATACCAAACATAAAAGAAAGCGTCTTTTGATAAAGTTG420              TTATTGGTGTAATGTGAAGGGACCACAATCATCACCATTCACCACTTGCTCCTAATTGAG480               TTGAAATCTTTTTACAACATAGAAAACTAGAAGATCGCCCTTTCTTGCTTCATATA TATA540              GATTTTGTATCATCGCAATTTCACATCACACACACAAGTAAAGCATTAGCAACCATAGCC600               ATGGCTTCCTCACCAAAACTCTTCTCTATGTCTATCCTCTTCCTTTTTGCCCTCTTCTCC660               ATGCAAATCCATGCTAGAGAGTACTTCAGCAAATTC CCAAGAGTTAACATCAATGAGAAA720              GAGACAACAACCAGAGAGCAAAAGCACGAGACCTTCGTTCCCCAGACCACCCAAAAGCCA780               GAAGAACAAGAGCCAAGGTTCATTCCTGAAACCCAAAATGGTTATGGCCTTTACGGCCAC840               GAGTCAGGCTCAAGCC GGCCCAGTTTCACCACCAAAGAAACCTATGAACCCTATGTCACC900              CCTGTTAGATTCCACCCTGATGAGCCCTATAACAGCATCCCCGAATCCTCCAACAATAAA960               GACACTTACTACTACAACAAGAATGCCTACGAGTCCACTAAGCAGCAAAACTTGGGCGAG102 0             GCCATTTTCACCGAGAAAGGATGGAGCACCAAGGAAAACCAGAACAACAACTACTACAAC1080              GGCAACAATGGTTACAACAATGGCGAGAAGCAAGGCATGAGCGATACTAGGTACTTGGAG1140              AATGGAAAGTACTACTATGACGTCAAGAGTGAGAACAACTATT ATCCAAACCGGTTCGAC1200             AACTCAAGAGGAGTTGCTTCGAGGAACGAGTTCAATGAGAATCGTTACAACAACATGGGA1260              AGGTACCACCAGAACCAAGAGGAGTTCGAGGAAAGCGAGGAAGAGTTCGAACCCTGATCA1320              CCTGTCGTACAGTATTTCTACAT TTGATGTGTGATTTGTGAAGAACATCAAACAAAACAA1380             GCACTGGCTTTAATATGATGATAAGTATTATGGTAATTAATTAATTGGCAAAAACAACAA1440              TGAAGCTAAAATTTTATTTATTGAGCCTTGCGGTTAATTTCTTGTGATGATCCGAATTCT1500              CGC CCTATAGTGAGTCGTATTAGTCGACGGTATCGATAAGCTTGATATCGAATTCCTGCA1560             GCCGGGGGATCCACTAGTTCTAGAGCGGCCGCCACCGCGGTGGAGCTCCAGCTTTTGT1618                (2) INFORMATION FOR SEQ ID NO:23:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1578 base pairs                                                    (B) TYPE: nucleic acid                                                       (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: DNA (genomic)                                             (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Gossypium hirsutum                                              (B) STRAIN: Coker 312                                                         (vii) IMMEDIATE SOURCE:                                                       (A) LIBRARY: Lambda DASH-CK                                                   (B) CLONE: CKE6-4A                                                            (x i) SEQUENCE DESCRIPTION: SEQ ID NO:23:                                     ATTTATATTAAATTAATATAGCATACTGGGTGAGAAGTAAAACTATTTAACAAATATATT60                CTTAAAAATACTAAAGGAGGTTTTGGTTGAATAGCAAAATATAAATATTACAAATTATAA120               AAATGTAGGTTCCAATATTTTTACTATTTTTCTATATA AAATGATAACCTTAAAAAGTAG180              TTTGTGGTTGATGGACTAATTTTTTAAAAAGAATTAATATTAGTAAAGATATATATGGTA240               CTAAACATAAGGAAATAGGGAAAACGTATCATATTTGTAGTGGGAGAATGGGATCATTAC300               AAGGAAAAATGAAATACA TATCCTTAACAACAAACATAAAAGAAAGCGTCTTTTGATAAA360              GTTGTTATTGGTGTAATGTGAAGGGACCACAATCATCACCATTCACCACTTGCTCCTAAT420               TGAGTTGAAATCTTTTTACAACATAGAAAACTAGAACATCTCCCTTTCTTGCTTCCTATA480               TATAGATTTTGTATCATCGCAATTCCACATCACACACGCAAGCAAAGCAAAGCATTAGCA540               ACCATAGCCATGGCTTCCTCACCAAAACTCTTCTCTATGTCTATCCTCTTCCTTTTTGCC600               CTCTTCTCCATGCAAATCCATGCTAGAGAGTACTTCAGCAAATTC CCAAGAGTTAACACC660              AATGAGAAAGAGACAACAACCAGAGAGCAAGAGCACGAGACCTTCGTTCCCCAGACCACC720               CAAAAGCCAGAAGAGCAAGAGCCAAGGTTCATCCCTGAAACCCAAAATGGTTATGGCCTT780               TACGGCCACGAGTCAGGCTCAGGCT CAGGCTCAGGCTCAAGCCGGCCCAGTTTCACCACC840              AAAGAAACCTATGAACCCTATGTCACCCCTGTTAGATTCCACCCTGATGAACCCTATAAC900               AGCATCCCCGAATCCTCCAACAATAAAGACACTTACTACTACAACAAGAATGCCTACAAG960               TCCAC TAAGCAGCAAAACTTGGGCGAGGCCATTTTCACCGAGAAAGGATGGAGCACCAAG1020             GAAAACCAGAACAACAACTACTACAACGGCAACATTAATGGCGAGAAGCAAGGCATGAGC1080              GATACTAGGTACTTGGAGAATGGAAAGTACTACTATGACGTCAAGAGTGAGA ACAGCTAT1140             TATCCAAACCAGCTCGACAACTCAAGAGGAGTTGCTTCCAGGAACGAGTTCGATGAGAAT1200              CGTTACAACAACATGGGAAGGTACCACCAGAACCAAGAGGAGTTCGAGGAAAGCGAGGAA1260              GAGTTCGAACCCTGATCACCTGTCGTACAGTA TTTCTACATTTGATGTGTGATTTGTGAA1320             GAACATCAAACAAAACAAGCACTGGCTTTAATATGATGATAAGTATTATGGTAATTAATT1380              AATTGGCAAAAACAACAATGAAGCTAAAATTTTATTTATTGAGCCTTGCGGTTACTTTCT1440              TGTGATGATCTT TTTTATTTTCTAATTATATATAGTTTCCTTCGCTTTGAAATGCTAAAG1500             GTTTGAGAGAGTTATGTTCTTTTTCTCTTCCTCTTTCTTTTTTAACTTTATCAAACAATT1560              TTTGAATAAAAATGTGAG 1578                                                       (2) INFORMATION FOR SEQ ID NO:24:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1618 base pairs                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: Genomic DNA                                               (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Cebia pentandra                                                 (vii) IMMEDIATE SOURCE:                                                        (A) LIBRARY: EMBL-CP                                                         (B) CLONE: E6-R5                                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:24:                                      TGGTATTTAGTATATTTAAATTTTAAATATTAATATATGTAAAATTAAAA50                          AAAAAAATTAGATTAGGATTTATTTTATAAAAAAAATGGAAATGAGATCA100                         TAAAAAGAGCACCAAATA ATAATAATAAAAGAAGAAATCAAAGTCAATCA150                        TTAACAACAAACACAAAGTGAAGAGGCCACTTTTGATAAAGTCTTATGTC200                         TCGTGCAAGGGACCACACACACAATCATCAGTTTTCACAGTCTCCCCCCC250                         GTCCCGTTTGCAACTAATTGAGTAGAAA ATTTTACAAATTGAGGGGAAAC300                        GAAAAAATTTGCCTTTCTATATAAACATTTCCTATCATCACAATTTCTCA350                         TTAGTGTGCACTCTCCCACGCAAAAAAAAAAAAAAAAAGAAAGAAAGCAT400                         TAGCTAGCCTTCCCCTTGCCCATGGCTTCCTCACCGAA ACTCGTTGCTAT450                        CTTCTTCCTCTTTGCCCTCTGCTCCATGCAGATTGATGCTAGAGAATTCT500                         TCAGCAAAGTCCCAAGTGTCAACACCAATGAGAAGGAGTCAACAACCATT550                         CCTGAGACCTTCATTCCCGTGACGACCACCCAAAAGACTTTGCTTCCC AA600                        CAAAGAAGAGCAGAGCACTTTCGGGAAGAACGAGCAAGAGCCAAGGTTTA650                         TCCCTGAGACTCAAAACGGATATGGCCTTTATGGTCACGAGTCAGGCCAG700                         CTCCCTCCCAGCACCACCACCAATACCAAAGAAACCTATGAACCCTATGT750                          TACCCCTGTTAGATTCCACCCTGATGAACCTTACAACAGCATTCCTGCAT800                        CCAAAACTAACAACAAAAATACTTACTATTACAACAAGAACCGCTATGAG850                         AATACCGAGAAACAAAATCTGGCTGAAGCCAGCTTCACAGAGAAAGGATG900                         GAGCACCAAG GAAAACCAGAACAACAACAACTACTACAACGGCAACAATG950                        GGTACAACAAGAATGCCTATGGGAATACCGAGCAGCAAAATTTGGGTGAG1000                        ACCATTTTCACAGAAAAAGGATGGAGCACCAAGGAAAACCAGAACAACAA1050                        CTACTATAATGGCAACAATG GATACAACAATGGTGAGAAGCAAGGCATGA1100                       GCGACACTAGATTCTTGGAGAATGGAAAGTACTACTATGATCTTAAGAAT1150                        GAGAACAACTACTATCCAAACCAGTTTGAGAACTCCAGGGGAGTTGCTTC1200                        AAGGAACGAGTTCAATGAGAATCGTTACAG CAACGTGGGAAGGTACAACC1250                       AGAACCAAGAGGAGTTCGAAGAGAACGAGGAAGAGTTCGAGCCATGAGCT1300                        AGCTGTCTTGTACTCTCTACAATGGAGTGAAAAACATCAAGCAAAACAAG1350                        AAGTGGTTTTAATTAGACGATAAGTATGCTATAATTAATT AGCAAAAACA1400                       GTAAAGAGAAAGATTTTATTTATTGGGTCTTGCGTTTAGTTTGTGATCTT1450                        TTCATTTTCTGGTTTGCATAGTATCCTTTGCTTTGCAATGCTCAAGATAT1500                        GAGAGTCATGCTTTTTATTTCTTTTCTACTTTATCAAACAATTTATTGAA 1550                       TAACAATGTAAGTATCTCCTAATAATCAGTCTTCAGTTTTCATATTCGCC1600                        TCTCTAGCAAATGCACCA1618                                                        (2) INFORMATION FOR SEQ ID NO:25:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1985 base pairs                                                   (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: double                                                     (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: DNA (genomic)                                             (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Gossypium barbadense                                            (B) STRAIN: Sea Island                                                        (vii) IMMEDIATE SOURCE:                                                       (A) LIBRARY: EMBL-SI                                                          (B) CLONE: SIH6                                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:25:                                      G TCGACCTGCAGGTCAACGGATCTTTTTTAGCTGTGTTTATTAAAAAAAA50                         TAAAAAAATATAAAAGTAGTTTTTTTAGAGTAAATGTAAAACTTTAAAAT100                         AATTGTAATATGTAAAATTAAAAATATTAAACTATTTACAACCGTCGGAT 150                        TAAAAATGATATATTTTTGAATGATGATGAAGATCGATTCCTGATGTATA200                         TAAATACTGCCTTCTATTCCCTTCAGTCTTCGCTTCACCCACTTTCTCAT250                         TTCACACGGGTTGTGGCGTAGTTTAAGCAGAGA GGGTGCGCAGGATAAAG300                        CTATTCACCATTGTTTCAACATGAAGGTTTGTAATAAAAATTTGTTTCTA350                         TCAGCATTGCTTTGCATTGCTGTTGCAGGAGTTTTGGGTCAAGCTCCTAG400                         TAATCCTCCTACGTCT ACGCCGGCGCCACCCACACCACCGGCTTCTACTC450                        CTCCTCCGACGACTCAAGCACCGCCTACACCAACCGCCACTCCGCCACCG500                         GTTTCTACTCCTCCTCCCACTTCATCACCGCCCCCAGTGACAGCTTCTCC550                         ACCCCCAGTTTCAACTCCTCCACCCAGTTCTCCTCCTCCTGCAACTCCAC600                         CACCTGCTTCTCCTCCTCCTGCAACTCCACCTCCAGCTTCTCCACCTCCT650                         GCCACTCCTCCACCAGCTTCTCCACCTCCCGCCACTCCACCACCTGCA AC700                        CCCACCGCCAGCAACTCCTCCTCCTGCTACCCCACCACCAGCTCCATTGG750                         CTTCTCCTCCAGCCACAGTCCCAGCTATCTCTCCAGTACAAACACCATTG800                         ACATCGCCACCAGCTCCGCCGACCGAGGCC CCAGCACCTACCCTCGGGGC850                        TGCTACGCCAGGTCCAGCTGGAACTGACACGGTACATTTTCTCTTATTCC900                         ACCATTTTATATCCTTCTTCTCCACCTACGATCAAGCTTTATTATCGGTT950                         GAAATTTAAGCCT TTACAGCAAGACTTAAAATATAATTTTATTAATGGTT1000                       TTATAATATTAAATTATAATTTTATCATTCTTACACATTAATATATAATG1050                        TGATAAAATTTTTTACTTTGGGTACCGTGCCAACTTCCTAACGTCGCCCT110 0                       TAATCTTACATAAACAACGATCTGAGCTTGTCTCGATATTAGCTAACCCT1150                        TAAGCCATTAGAGATGGCTATTGGTTCCGTCTCGACATACCTTCAACATA1200                        ATCTGATTTAAAATTAAATACTTATATCTATTTTTTAACACAATA TTTAA1250                       AATTATCTATAATTTCTTCTCAACTTATAATTAAAATAACAATACTTCAG1300                        CGTATTCAAATTTACGTACCTATGTTAATCTAATTTGACAATAATATTTA1350                        TGTTAATTGAATTAAAGCTTGAGATATT AAATTTAATTAAGACTCAACAT1400                       TGACGACAGTACGATTCAACGTATTAGATTTAATTAATGTTGGATTTTGA1450                        ACCTATTATTGCAGAGTGGAGCAAATCAAATGTGGACCGTACAAAAGATG1500                        ATGGGAAGCT TAGCCATGGGATGGGCTCTGCTCAATCTGATGGTTTAAAA1550                       CAAAAGAGTGCCTCACATTTGATGCAATAGCTCTGTAATGTTTCATTCAT1600                        TTGCTTATTTCGGCCTTGTTTTTCTCGTATTCTATGGGCTGATGTCTCAT 1650                       ATGGGACTTTTCTACTAGAGAGCCTACGTTACTTTACCATTATATTGTAT1700                        TCTTTGAGACATTATTATTATTTTTTTACCTTTTGAGGACACTCTTTTTT1750                        TGTATTTGAAGGAATTTATTGTTTATTTTGTTTGGAATATGT TTGGTTGG1800                       ATTTATTCGATTCATATATATTATATAAAAGTAATTATGTTATTAAGAAA1850                        CGTAGTAAGAACTTACAAATATAAGGATCGAATCCCGAACTTCATGCAAA1900                        TCAATTTACAACCCACACAAGTTTA ACATTAAATTAACGTGATTGGTTAG1950                       TAAATTCATGTTTCTCTGTTTAATTTGTTGAATTC1985                                       (2) INFORMATION FOR SEQ ID NO:26:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 2415 base pairs                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: DNA (genomic)                                             (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Gossypium barbadense                                            (B) STRAIN: Sea Island                                                        (vii) IMMEDIATE SOURCE:                                                       (A) LIBRARY: EMBL-SI                                                          (B) CLONE: SIB12                                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:26:                                      CTGGCCTTTTGCTGG CCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCC50                         TGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTC100                         GCCGCAGCCGAACGACCGAGCGCACGGGNTCAGTGAGCGAGGAAGCGGAA1 50                        GAGCGAAAAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTA200                         ATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCA250                         ACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGG CTTTACAC300                        TTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATT350                         TCACACAGGAAACAGCTATGACCATGATTACGCCAAGCTCGAAATTAACC400                         CTCACTAAAGGGAACAAAAGCT GGAGCTCCACATGGTTTAATTAAACATT450                        ATGTTCCATCCATCTATATTTATATCCATTAAAACAAGTCGTTGAGCAAA500                         TAATGGATACTGGATACCATCATATCTATGATTAAAATTTTGCATGTGCC550                         CT TTTAATGTATAGCTTAATTATCCTCCAAATTTGTACTCTTTCACCACT600                        AATTAGCTACGTACGTTACTTAGCTTTGCTTGTCGTCATCTTCTGTACTA650                         CAAACTCTTTCTCTTTTTGTATAAATAGCTATACACTTTTTCTCTCCTCA 700                        AATCAATAAGGTTAGGTCAACCAATTGTTTGAGCTAGCTAGCTCTTACTC750                         AAATGGCAACCAAAACGATGATGTTGCAAATATTTTCACTTTTCTTCTTT800                         TTGTTCAGTGTATGCAACTCCATTTTCCTT GGTGCTAATGGAGATGACAA850                        TGGTGGTTGCAAACTGCCCATGCACCTTCTACGGTGGTGCTGATGCTACC900                         GGCACAATGGGTGAGTTTCAAACTTTCAAACCATTACCTACATAAAAATC950                         TCTAGGCTAT GTTCTTAATTTGTGATGTTTCTATAGGGGGAGCTTGTGGT1000                       TATGGAAACCTGTACAGTCAAGGGTATGGAACGAGCACAGCAGCTTTGAG1050                        CACTGCACTTTTCAACAATGGCTTGAGCTGCGGTGCACTGCTACGAGCTC 1100                       CGGTGCAACAATGATCCTCAATGGTGCATTAGTCGAACCATAACCGTGAC1150                        AGCCACCAACTTTTGTCCCCCTAACTATGCTTTATCTAGTGACAATGGCG1200                        GGTGGTGCAATCCCCCACGAGAACACTTTGATTTGGC CGAACCGGCATTC1250                       TTGCAGATCGCGGAATATCGAGCTGGAATCGTCCCTGTTATGTTCAGAAG1300                        GTGGTGAATAAAACTCAATTCAAATCATCACACTCTTTAAGGTATGTTAA1350                        ACTGTTGGGTGTTTAAC CTTTTGCAGGGTGTCATGTGTGAAGAAAGGAGG1400                       CATCAGGTACACCATGAATGGACATTCGTACTTCAACATGGTGTTGATAA1450                        CCAACGTGGGAGGGGCAGGGGATATAACGTCAGTGTCCATCAAGTGTTCC1500                        AAAACAGGATGGCTACCTATGTCCAGAAATTGGGGCCAAAACTGGCAGAG1550                        CAATGCTTACCTTAACGGCCAAAGCCTCTCTTTCAAAGTGACTGCCAGCG1600                        ATGGCAGGACTATCACAAACTACAATGTAGTGCCTGCTGGTTGG CAATTC1650                       GGACAAACTTTTGAAGGAGGCCAGTTTTAAGACAATATTATAGTGTCTGT1700                        CTAATATTAAAACTGGAATTGACATATTACTTATATAAGGCACATGAGCG1750                        TTTTATGCCGAGGTAGTAAAGTGG CGCCCGCTGCGTTTATGTGTGAAATA1800                       GGCGAGCAAGTGCCATTAGCCTATAATATATACATTTCTTATAGTGAACC1850                        AAACTATTAAGTTTGAACTCTAGAGGTGATATCCATAATGTCTGAAATTT1900                        GATT GTTGATGATTGACCATGATATTTATGGTTTTCATTATTGAAATACT1950                       TTTTTTTTATAATTTATAAATAATGGGTCATTTCTTTTTACAAATATTTT2000                        CGACATATTTTATGATTTGTCAGCTAAATATTATTAATCAAAATTAGGAT 2050                       GCAATATTGAATCAAGAATGTATAAATGAATTATTGAGACATCATATAAG2100                        ATATAAAAGATGTATCATATTTTTTACGTTGAGCAGTCATACAATAAAAT2150                        TAATCTCCTAATATAAGATAGTATTTCAGTA GTGCATATGTTGAATGGTA2200                       ACTTTGTTGTGAGGCAAATAATTTTGCCCAAGGTGCTATGTAGCAGAGAG2250                        CGGGAGTTGCACATTTGTCATAATTTAGGGAGCCAAACCATGCAACGGAT2300                        TTCATGGCCAA GATAGTCATTGGGGTCGGGACTCCCAAAATTTTAATTAG2350                       TCCCTCCTTCAAGTTTTTCTAAATTTTCAAATATTTTAATTGGACTCCTC2400                        TAACTTTTCTTTTTT 2415                                                          (2) INFORMATION FOR SEQ ID NO:27:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 279 base pairs                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: DNA (genomic)                                             (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Gossypium barbadense                                            (B ) STRAIN: Sea Island                                                       (vii) IMMEDIATE SOURCE:                                                       (A) LIBRARY: EMBL-SI                                                          (B) CLONE: SIA-11-B                                                           (xi) SEQUENCE DESCRIPTION: SEQ ID NO:27:                                      CCCTTTTCCCAACCCTTTTCTCCTGTATCAAAGACTTGGTGGTGCAACAAGCCTTGTTCT60                CAACCTTGGAGGATTGGCTAGTGAAGCATCCCAAAATCCTCCAGTTT TCATGGGAAAATG120              GCCAAACACCAGCCTCCTCTCACCGCTTCCTCACCCTCACTGTCTTGTCTTACATCTCTT180               TCACATTCGTTCTCTCCCAACTATCTCGCCCTTCACTTTCACGTCCACTCCTCAAATCAA240               TCGCAGCTGTCCACAACATCTTCCTCC TTACCCTTTCTT279                                   (2) INFORMATION FOR SEQ ID NO:28:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 2539 base pairs                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: DNA (genomic)                                             (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Gossypium barbadense                                            (B) STRAIN: Sea Island                                                        (vii) IMMEDIATE SOURCE:                                                       (A) LIBRARY: EMBL-SI                                                          (B) CLONE: SIB8                                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:28:                                      TTTTTAATGGTGTTGGATGGTTATATTATATCTCGATTATATATATTTTTTTTAAAAACC60                GAAGTTGAATGTCTA AATAGGAAGTAATTTTTTTAATATTATTTTTTTATAATATTTGAA120              TCCGATATCTTATTTAAAAACCATCGAAATTTTTATTACTCAATCATTACCGAAATAGAA180               TCGGGCTAAAATATTTCGAAAACTAAAAGTTTCACTTTTTATATTGAAAAACGAGGCTTT2 40              GTGATTCTTATAAATTTAATTCATTGAAATTTCATCAAGTAAAACAGAAGAATTATAAAT300               CTCTAAAATGATAGATAAAAATGTCGCAAATAAAGCCATTGTGACACCCATTAAAGGAGT360               CTTTTCCCATCCAGGGGCTACCTTACCATATTCCGAATTCAA TGGTTTCAAAAAATCTCC420              TACAACAATTCGTCTTGGACTAGATCTAGAACTACTGTTAACGTTTTGTGTAGCCATAAA480               TCTTATTTTGTATTCATTGAGATTTGTTAACTTTGTATATCATTTCACTATAAATAAACA540               ATCTTATCATAGACCCATTGTG ATCATGAAATTATATAATAATGGAAACAAAAACCTTAG600              TTGCAATCTCTATATCTGATTTACTTGTAAGTTTTACTAGGTACGCCTTATATACTGCTT660               TTGGGTAACTCTCTCAACAATTAAGAGATCCATTCGAGGAATACGGGAAATAATTGAAGA720               AA CGAGCCTCCCCATATGTATGTCTATTTGGTCACATATCCACTCCAAAATCTACTCTGT780              TTTCCCTGCTTTTCCAAGCAATGTAGTTGCTCATCAATTTCTTTCCTTTCAAATAAACAT840               CCCAACAGAGTTTGCTAAAGTTGATAAGTATGTCTTCTAGTTAACTAAAG TAGCATATTT900              TACCTAACTTCACCCTCCAATATCCTAAATAAAAAGCTCCCATTTCTTATCCAATCAAAA960               CATCCATAACATTTTTGTTCAAGGACCACTTCTTCCCTTCCATTTTACTTTGTTTTAGTT1020              GCCATAACGTCACCTTCCAATACAACCCAC AATGAGGCAACAATATGTCTTCACTACTCT1080             CACGTTGCTCATCCTGTTTTCCCTCAGTTGTTCAACAACATTAGCCCAATCTCCGGCACT1140              GGCCCCGGCACCTTCTGGTCCGACAAACGTCACCAAGATCCTCGAAAAAGCTGGTCAATT1200              CACCCTCTTC ATTCGTCTTCTAAAGTCCACTCAAGTGGCCAACCAGCTGCTCGGTCAGCT1260             CAACAATTCCAACAATGGTATGACCGTTTTTGCACCAACGGACAATGCTTTCTCCAGCCT1320              TAAATCGGGCACATTGAATTCACTCACCGATGAACAAAAAGTGCAGCTGGTGCAATT CCA1380             CATCGTCCCAACATACCTCACCTCGTCTCAGTTCCAAACCATTAGCAACCCTTTGAGAAC1440              CCAAGCTGGTGATAGTGGCGATGGCAAGTTCCCTCTCAATGTCACCACTTCGGGGAACTC1500              TGTGAATATAACAACAGGGTTGACAAACACCAGTGTT TCCGGCACTATTTACACTGATGG1560             TCAGCTTGCTGTTTATCAAATCGATCAAGTTCTTCAACCATTGCAAATATTTGCACCTAG1620              GCCTCCAGCTCCCGCACCGGCACCGGCAAAGTCGAAGAATAAGAAGGCTACCACTGTTGC1680              TGATAGCCCCGATGTTA CCCCAGCTGATAACTCCAAAGCGGCCACCTTGCAAAATGCTGG1740             TTTGTTTGGAGTTGCTGCTCTAGTTATTGCACTTTCTTTGTGACCATGAAAATGGAGAAA1800              AGAAGAAGACAGTGATTTTGATGGTGGTGATCAAATTGGAGTAAATAGTGAAATTAAAAA1860              TAGATATAATTGAGTTATTTTGTAAACATATTAAATTCCTTATTTATTATTTATATACTT1920              ACTTACTAAGCTATAAGCTTACTTTCTTTTCTCTCTTTTGTTTTATAGTGGCATCCAGCT1980              AGCACAGGAGTTAGAGATCGTTGAAGATCTGCTCGCACTATCAA TACTTTTGGTATTTGA2040             ACTCAACATTTTGAATTATGGCATGTATAGTAGGCTTAGTTATTTTGTTACGTGTCATAA2100              GTGCCTAGGTTTAAAATATCGGTTATAGTATTTGACCGATTATTTTGTACAAAGCCATTG2160              AAGTTGGCTAATATTGTTAAAGGC ATATGTTATAACGATTCATGATCTAATTGAATGTCT2220             CGGTTTTATTTAATGGTATGTGTTATATTCTGGTAATACCTCGTACCCTGTTTCAGTGTT2280              GAATGCGAGTAAGGGGCGTTACAATAGCCTTCATAAACAAGTTCACCAACGGCTTGAAAT2340              ACTC TAAAAGGTAAGGATAAGAACAAGAACTCAATCTTTATTCAATACACTCATGGTTTT2400             ATCAAGAAGATCGGTTGAAGAGTTAGAAAGTCCTAGTTGCACATCACATGTTATTTTGTC2460              AAACATGTTACTACTTTAAAAGCCGAAAACATTGACTGTCCATGAGGTCGA AACCTTCCA2520             CACAGTTGAAACAAAAAAA2539                                                       (2) INFORMATION FOR SEQ ID NO:29:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 562 base pairs                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                          (i i) MOLECULE TYPE: DNA (genomic)                                            (iii) HYPOTHETICAL: NO                                                        (iv) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Gossypium barbadense                                            (B) STRAIN: Sea Island                                                        (vii) IMMEDIATE SOURCE:                                                       (A) LIBRARY: EMBL-SI                                                          (B) CLONE: SIB6                                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:29:                                      TTTTTCAAATCGAATCGAGTTTTGCTCACCCTAAGACAAC CATATCTAATCCAATATCTG60               GTTTTCAAATATTTTCTTTTAATCATGGTTAGTTTTTTTGTTAATTTTCTATACCTACTT120               TTTACCTTAAACTTCACATCCATTTTGATCACCAACAACTGGTACAACCCCACACGTTGT180               TTTTTTTCTGTTTATATCTT ACAACTTACAACCGAAACATCCACAGTACACACACATATA240              TATATTCATCCAATCTGCTAAATTGGTATCCAAAACTATTCAACTTTCTCTCTGAACTCC300               TCCAAGTTAGGTTAGTGTTTTCCATAATTTGCATTTGTGTTAAAAGTTGCTTCTCTTGAG360                AGTTCAAAGGATTCATTTTTCTTTCAAGTTACATGCATGTCTATGTTTTGAAATGGGGTT420              TACTTTTTTCTTTTGTTCATAATGTAAATTTATTGGATATTTTGCTGTTTATCTGCAAGT480               AATTGCCAATGATTTGATTCTTGTAGGAGTTTGGAGAGTGGTCACCG CTGAAGCAAAGCC540              AAATTTCTTGGGGGAAAAGAAA562                                                     (2) INFORMATION FOR SEQ ID NO:30:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 18 base pairs                                                     (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                           (ii) MOLECULE TYPE: Oligonucleotide                                          (iii) HYPOTHETICAL: YES                                                       (xi) SEQUENCE DESCRIPTION: SEQ ID NO:30:                                      ACCGAAATAGAATCGGGC18                                                          (2) INFORMATION FOR SEQ ID NO:31:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 35 base pairs                                                     (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: Oligonucleotide                                           (iii) HYPOTHETICAL: YES                                                       (xi) SEQUENCE DESCRIPTION: SEQ ID NO:31:                                      CGCGGTTCGAACCATGGTTTAGAGATTTATAATTC35                                         (2) INFORMATION FOR SEQ ID NO:32:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 21 base pairs                                                     (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: Oligonucleotide                                           (iii) HYPOTHETICAL: YES                                                       (xi) SEQUENCE DESCRIPTION: SEQ ID NO:32:                                      GGAGAATGCTCGGGAGGTGGT21                                                       (2) INFORMATION FOR SEQ ID NO:33:                                             (i) SEQUENCE CHARACTERISTICS:                                                 ( A) LENGTH: 35 base pairs                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: Oligonucleotide                                           (iii) HYPOTHETICAL: YES                                                       (xi) SEQUENCE DESCRIPTION: SEQ ID NO:33:                                      TAGATCTCGAGCCATGGTAACACAACAAGCCTTTT35                                     

I claim:
 1. A method of obtaining a DNA sequence which preferentiallypromotes gene expression in fiber cells comprising the steps ofa)screening genomic DNA fragments obtained from a fiber-producing plantwith a nucleotide sequence selected from either strand of the groupconsisting of SEQ ID NOS: 2, 5 and 7 and the first 541 bp of SEQ IDNO:20; the first 581 bP of SEQ ID NO:21, the first 567 bp of SEQ IDNO:22, the first 512 bp of SEQ ID NO:23, the first 421 bp of SEQ IDNO:24; the first 250 bp of SEQ ID NO:25; the first 279 bp of SEQ IDNO:27; and the first 287 bp of SEQ ID NO:28, and selecting those genomicfragments which hybridize with said either strand; b) isolating aputative promoter sequence from said selected genomic fragments, and c)testing the putative promoter sequence for efficacy as a fiber-specificpromoter by operably joining the putative promoter to a reporter genecoding sequence to form an expression construct and obtaining a whole,fertile transgenic cotton plant containing said expression construct sothat expression of the reporter gene can be observed in the cells of theplant; and (d) selecting those promoter sequences of step (b) whichpreferentially express the reporter gene in the fiber cells of the plantof step (c).